T-cell modulatory multimeric polypeptides and methods of use thereof

Abstract

The present disclosure provides T-cell modulatory multimeric polypeptides (TMMPs) that comprise comprise (i) an optional immunomodulatory polypeptide such as a variant IL-2 polypeptide, (ii) class I HLA major histocompatibility complex (MHC) polypeptides (a class I HLA heavy chain polypeptide and a β2 microglobulin polypeptide), (iii) a peptide that presents an epitope to a T-cell receptor, which together with the class I MHC polypeptides forms a peptide-MHC complex (pMHC), (iv) a tumor-targeting polypeptide, and (v) an optional Ig Fc polypeptide or other scaffold. Such TMMP is useful for modulating the activity of a T cell, and for modulating an immune response in an individual, and for "redirecting" a patient's repertoire of antiviral T cells to attack and kill cancer cells.

Description

T-CELL MODULATORY MULTIMERIC POLYPEPTIDES AND METHODS OF USE THEREOF CROSS-REFERENCE

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 63 / 283,028, filed November 24, 2021, U.S. Provisional Patent Application No.63 / 309,288, filed February 11, 2022, and U.S. Provisional Patent Application No.63 / 338,131, filed May 4, 2022, which applications are incorporated herein by reference in their entirety. INCORPORATION-BY-REFERENCE OF MATERIAL ELECTRONICALLY SUBMITTED

[0002] A Sequence Listing is provided herewith as a Sequence Listing XML, “CUEB- 145WO_SEQUENCE_LIST” created on November 17, 2022 and having a size of 1,190,000 bytes. The contents of the Sequence Listing XML are incorporated by reference herein in their entirety. INTRODUCTION

[0003] An adaptive immune response involves the engagement of the T cell receptor (TCR), present on the surface of a T cell, with a small peptide antigen non-covalently presented on the surface of an antigen presenting cell (APC) by a major histocompatibility complex (MHC; also referred to in humans as a human leukocyte antigen (HLA) complex). This engagement represents the immune system’s targeting mechanism and is a requisite molecular interaction for T cell modulation (activation or inhibition) and effector function. Following epitope-specific cell targeting, the targeted T cells are activated through engagement of costimulatory proteins found on the APC with counterpart costimulatory proteins the T cells. Both signals – epitope / TCR binding and engagement of APC costimulatory proteins with T cell costimulatory proteins – are required to drive T cell specificity and activation or inhibition. The TCR is specific for a given epitope; however, the costimulatory protein not epitope specific and instead is generally expressed on all T cells or on large T cell subsets. SUMMARY

[0004] The present disclosure provides T-cell modulatory multimeric polypeptides (TMMPs) that comprise an immunomodulatory polypeptide, class I HLA polypeptides (a class I HLA heavy chain polypeptide and a β2 microglobulin polypeptide), a peptide that presents an epitope to a T-cell receptor, and a tumor-targeting polypeptide. A TMMP of the present disclosure is useful for modulating the activity of a T cell, and for modulating an immune response in an individual and for “redirecting” a patient’s repertoire of antiviral T cells to attack and kill cancer cells.BRIEF DESCRIPTION OF THE DRAWINGS

[0005] FIG.1A-1B provide an amino acid sequence of a wild-type human β2M polypeptide (SEQ ID NO:1) (FIG.1A) and an amino acid sequence of a β2M polypeptide with an R12C substitution (SEQ ID NO:2) (FIG.1B).

[0006] FIGs.2A-2M provide amino acid sequences of immunoglobulin Fc polypeptides (SEQ ID Nos: 3-15, respectively).

[0007] FIG.3A-3E provide amino acid sequences of wild-type HLA-A*0201 (SEQ ID NO:16) (FIG. 3A) and variants (SEQ ID Nos: 17-20, respectively) (FIG.3B-3E).

[0008] FIG.4A-4E provide amino acid sequences of wild-type HLA-A*1101 (SEQ ID NO:21) (FIG. 4A) and variants (SEQ ID Nos: 22-25, respectively) (FIG.4B-4E).

[0009] FIG.5A-5E provide amino acid sequences of wild-type HLA-A*2402 (SEQ ID NO:26) (FIG. 5A) and variants (SEQ ID Nos: 27-30, respectively) (FIG.5B-5E).

[0010] FIG.6A-6E provide amino acid sequences of wild-type HLA-A*3303 (SEQ ID NO:31) (FIG. 6A) and variants (SEQ ID Nos: 32-35, respectively) (FIG.6B-6E).

[0011] FIGs.7A-7B provide an alignment of HLA-A heavy chain amino acid sequences (FIG.7A; SEQ ID NOs:36-44, respectively) and a consensus sequence (FIG. 7B; SEQ ID NO: 45).

[0012] FIGs.8A-8B provide an alignment of HLA-B heavy chain amino acid sequences (FIG.8A; SEQ ID NOs:46-52, respectively) and a consensus sequence (FIG. 8B; SEQ ID NO: 53).

[0013] FIGs.9A-9B provide an alignment of HLA-C heavy chain amino acid sequences (FIG.9A; SEQ ID NOs:54-61, respectively) and a consensus sequence (FIG. 9B; SEQ ID NO: 62).

[0014] FIG.10 provides a consensus amino acid sequence for each of HLA-E, -F, and -G heavy chains (SEQ ID NOs: 63-65, respectively). Variable amino acid (aa) positions are indicated as “X” residues sequentially numbered; the locations of amino acids 84, 139, and 236 are double underlined.

[0015] FIG.11 provides an alignment of consensus amino acid sequences for HLA-A (SEQ ID No:45), -B (SEQ ID NO:53), -C (SEQ ID NO:62), -E (SEQ ID NO:63), -F (SEQ ID NO:64), and -G (SEQ ID NO:65).

[0016] FIGs.12A-12D provide amino acid sequences of a wild-type IL-2 polypeptide (SEQ ID NO:66) (FIG.12A), IL-2Rα (SEQ ID NO:67) (FIG.12B), IL-2Rβ (SEQ ID NO:68) (FIG.12C), and IL-2Rγ (SEQ ID NO:69) (FIG.12D).

[0017] FIG.13A-13K provide amino acid sequences of SARS-CoV-2-encoded polypeptides (SEQ ID Nos:70-94).

[0018] FIG.14 provides CD8 T cell epitopes of SARS-CoV-2-encoded polypeptides and primary HLA class I heavy chain allele restriction.

[0019] FIG.15 provides CD8 T cell epitopes of SARS-CoV-2-encoded polypeptides and HLA class I heavy chain allele restriction.

[0020] FIG.16A-16D provide amino acid sequences of HLA-E heavy chains (SEQ ID Nos:837-840, respectively).

[0021] FIG.17A-17D provide amino acid sequences of HLA-G heavy chains (SEQ ID Nos:841-844, respectively).

[0022] FIG.18A-18D provide schematic depictions of TMMPs.

[0023] FIG.19 provides amino acid sequences of SARS-CoV-2 peptides and HLA class I heavy chain allele restriction.

[0024] FIG.20A-20J provide amino acid sequences (FIG. 20A (SEQ ID NO:867), 20C (SEQ ID NO:868), 20E (SEQ ID NO:869), 20G (SEQ ID NO:870), and 20I (SEQ ID NO:871)) and descriptions (FIG.20B, 20D, 20F, 20H, and 20J) of exemplary TMMP “heavy chain” constructs.

[0025] FIG.21A-21E provide amino acid sequences and descriptions of exemplary TMMP “light chain” constructs.

[0026] FIG.22A-22H provide amino acid sequences of exemplary anti-mesothelin scFv.

[0027] FIG.23A-23D provide amino acid sequences of exemplary anti-TROP-2 scFv.

[0028] FIG.24A-24B provide amino acid sequences of exemplary “mAb fusion” constructs.

[0029] FIG.25A-25B provide amino acid sequences of exemplary “mAb fusion” constructs.

[0030] FIG.26A-26B provide amino acid sequences of exemplary “mAb fusion” constructs.

[0031] FIG.27A-27B provide amino acid sequences of exemplary anti-CD28 scFv.

[0032] FIG.28A-28C provide schematic depictions of TMMPs, showing various possible MOD positions and various possible TTP positions.

[0033] FIG.29 depicts PBMC reactivity to the SARS-CoV-2 HLA-A*0201 peptide YLQPRTFLL (SEQ ID NO:218).

[0034] FIG.30 depicts expansion of SARS-CoV-2 (SCV2)-reactive CD8+T cells.

[0035] FIG.31A-31D depict the effect of various constructs on activation of SARS-CoV2-specific T cells present in a population of PBMCs (FIG.31A and FIG.31B) and PBMC-mediated killing of target B cells (FIG.31C and FIG. 31D).

[0036] FIG.32A-32G provide amino acid sequences of polypeptide chains of an exemplary TMMP, where FIG.32A provides the amino acid sequence of the polypeptide designated “4518” (SEQ ID NO:867), FIG.32B provides the amino acid sequence of the polypeptide designated “4700” (SEQ ID NO:883), and FIG.32C-32G provide amino acid sequences of components of the 4518 and 4700 polypeptides.

[0037] FIG.33 provides the amino acid sequence of the polypeptide designated “4656” (SEQ ID NO:910)

[0038] FIG.34 provides the amino acid sequence of the polypeptide designated “3534” (SEQ ID NO:911).

[0039] FIG.35 provides the amino acid sequence of the polypeptide designated “1717” (SEQ ID NO:912).

[0040] FIG.36A-36B depict the effect of various constructs on expansion of SARS-CoV2-specific CD8+T cells (FIG.36A) or CMV-specific CD8+T cells present in a population of PBMCs.

[0041] FIG.37A-37B depict the effect of various constructs on killing of CD19+tumor cells.

[0042] FIG.38A-38L provide amino acid sequences of various polypeptides suitable for including in a TMMP (FIG. 38A-38H) (SEQ ID Nos: 1209-1216, respectively) and certain components of the polypeptides (FIG.38I-38L) (SEQ ID NOs: 872, 18, 877, and 878 respectively).

[0043] FIG.39A-39B provide examples of amino acid sequences of polypeptides suitable for including in a TMMP (SEQ ID Nos: 1217 and 1218, respectively).

[0044] FIG.40A-40B provide examples of amino acid sequences of polypeptides suitable for including in a TMMP (SEQ ID Nos: 1219 and 1218, respectively).

[0045] FIG.41A-41C provide examples of amino acid sequences of polypeptides suitable for including in a TMMP (SEQ ID Nos: 870, 1220 and 883, respectively).

[0046] FIG.42A-42C provide examples of amino acid sequences of polypeptides suitable for including in a TMMP (SEQ ID Nos: 1221-1223, respectively).

[0047] FIG.43A and 43B provide amino acid sequences of constructs 1715 (FIG.43A) (SEQ ID NO:1224) and 1717 (FIG.43B) (SEQ ID NO:912).

[0048] FIG.44 depicts the effect of linker length on TMMP-mediated target tumor cell killing.

[0049] FIG.45 depicts the effect of copy number of variant IL-2 polypeptide on TMMP-mediated target tumor cell killing.

[0050] FIG.46 depicts TMMP-mediated anti-viral T cell expansion. DEFINITIONS

[0051] The terms “polynucleotide” and “nucleic acid,” used interchangeably herein, refer to a polymeric form of nucleotides of any length, either ribonucleotides or deoxyribonucleotides. Thus, this term includes, but is not limited to, single-, double-, or multi-stranded DNA or RNA, genomic DNA, cDNA, DNA-RNA hybrids, or a polymer comprising purine and pyrimidine bases or other natural, chemically or biochemically modified, non-natural, or derivatized nucleotide bases.

[0052] The terms ''peptide," ''polypeptide," and "protein" are used interchangeably herein, and refer to a polymeric form of amino acids of any length, which can include coded and non-coded amino acids, chemically or biochemically modified or derivatized amino acids, and polypeptides having modified peptide backbones. Furthermore, as used herein, a "polypeptide" refers to a protein that includes modifications, such as deletions, additions, and substitutions (generally conservative in nature as would be known to a person in the art) to the native sequence, as long as the protein maintains the desired activity. These modifications can be deliberate, as through site-directed mutagenesis, or can be accidental, such as through mutations of hosts that produce the proteins, or errors due to polymerase chain reaction (PCR) amplification or other recombinant DNA methods. References herein to a specific residue or residue number in a known polypeptide are understood to refer to the amino acid at that position in the wild-type polypeptide. To the extent that the sequence of the wild-type polypeptide is altered, either by addition or deletion of one or more amino acids, one of ordinary skill will understand that a reference to the specific residue or residue number will be correspondingly altered so as to refer to the same specific amino acid in the altered polypeptide, which would be understood to reside at an altered position number. For example, if an MHC class I polypeptide is altered by the addition of one amino acid at the N-terminus, then a reference to position 84 or a specific residue at position 84, will be understood to indicate the amino acids that are at position 85 on the altered polypeptide. Likewise, a reference herein to substitution of a specific amino acid at a specific position, e.g., Y84, is understood to refer to a substitution of an amino acid for the amino acid at position 84 in the wild-type polypeptide. A Y84C substitution is thus understood to be a substitution of Cys residue for the Tyr residue that is present in the wild-type sequence. If, e.g., the wild-type polypeptide is altered to change the amino acid at position 84 from its wild-type amino acid to an alternate amino acid, then the substitution for the amino acid at position 84 will be understood to refer to the substitution for the alternate amino acid. If in such case the polypeptide is also altered by the addition or deletion of one or more amino acids, then the reference to the substitution will be understood to refer to the substitution for the alternate amino acid at the altered position number. A reference to a “non-naturally occurring Cys residue” in a polypeptide, e.g., an MHC class I polypeptide, means that the polypeptide comprises a Cys residue in a location where there is no Cys in the corresponding wild-type polypeptide. This can be accomplished through routine protein engineering in which a cysteine is substituted for the amino acid that occurs in the wild- type sequence.

[0053] A polynucleotide or polypeptide has a certain percent "sequence identity" to another polynucleotide or polypeptide, meaning that, when aligned, that percentage of bases or amino acids are the same, and in the same relative position, when comparing the two sequences. Sequence identity can be determined in a number of different ways. To determine sequence identity, sequences can be aligned using various convenient methods and computer programs (e.g., BLAST, T-COFFEE, MUSCLE,MAFFT, etc.), available over the world wide web at sites including ncbi.nlm.nili.gov / BLAST, ebi.ac.uk / Tools / msa / tcoffee / , ebi.ac.uk / Tools / msa / muscle / , mafft.cbrc.jp / alignment / software / . See, e.g., Altschul et al. (1990), J. Mol. Bioi.215:403-10. Unless otherwise stated, “sequence identity” as referred to herein is determined by BLAST (Basic Local Alignment Search Tool), as described in Altschul et al. (1990) J. Mol. Biol.215:403.

[0054] The term "conservative amino acid substitution" refers to the interchangeability in proteins of amino acid residues having similar side chains. For example, a group of amino acids having aliphatic side chains consists of glycine, alanine, valine, leucine, and isoleucine; a group of amino acids having aliphatic-hydroxyl side chains consists of serine and threonine; a group of amino acids having amide containing side chains consisting of asparagine and glutamine; a group of amino acids having aromatic side chains consists of phenylalanine, tyrosine, and tryptophan; a group of amino acids having basic side chains consists of lysine, arginine, and histidine; a group of amino acids having acidic side chains consists of glutamate and aspartate; and a group of amino acids having sulfur containing side chains consists of cysteine and methionine. Exemplary conservative amino acid substitution groups are: valine- leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine-glycine, and asparagine- glutamine.

[0055] The term “immunological synapse” or “immune synapse” as used herein generally refers to the natural interface between two interacting immune cells of an adaptive immune response including, e.g., the interface between an antigen-presenting cell (APC) or target cell and an effector cell, e.g., a lymphocyte, an effector T cell, a natural killer cell, and the like. An immunological synapse between an APC and a T cell is generally initiated by the interaction of a T cell antigen receptor and major histocompatibility complex molecules, e.g., as described in Bromley et al., Annu Rev Immunol. (2001) 19:375-96; the disclosure of which is incorporated herein by reference in its entirety.

[0056] “T cell” includes all types of immune cells expressing CD3, including T-helper cells (CD4+cells), cytotoxic T-cells (CD8+cells), T-regulatory cells (Treg), and NK-T cells.

[0057] The term “immunomodulatory polypeptide” (also referred to herein as a “MOD”), as used herein, means a polypeptide that specifically binds a cognate costimulatory polypeptide on a T cell, thereby providing a signal which, in addition to the primary signal provided by, for instance, binding of a TCR / CD3 complex with a major histocompatibility complex (MHC) polypeptide loaded with peptide, mediates a T cell response, including, but not limited to, proliferation, activation, differentiation, and the like. As discussed herein, a MOD can include, but is not limited to wild-type or variants of wild-type polypeptides such as a cytokine (e.g., IL-2), CD7, B7-1 (CD80), B7-2 (CD86), PD-L1, PD-L2, 4-1BBL, OX40L, Fas ligand (FasL), inducible costimulatory ligand (ICOS-L), intercellular adhesion molecule (ICAM), CD30L, CD40, CD70, CD83, HLA-G, MICA, MICB, HVEM, lymphotoxin beta receptor, 3 / TR6, ILT3, ILT4, HVEM, an agonist or antibody that binds Toll ligand receptor, and a ligand thatspecifically binds with B7-H3. A MOD of a TMMP can bind a cognate costimulatory polypeptide (i.e., a “co-MOD”) that is present on a target T cell.

[0058] As used herein the term “in vivo” refers to any process or procedure occurring inside of the body.

[0059] As used herein, "in vitro” refers to any process or procedure occurring outside of the body.

[0060] “Heterologous,” as used herein, means a nucleotide or polypeptide that is not found in the native nucleic acid or protein, respectively.

[0061] "Recombinant," as used herein, means that a particular nucleic acid (DNA or RNA) is the product of various combinations of cloning, restriction, polymerase chain reaction (PCR) and / or ligation steps resulting in a construct having a structural coding or non-coding sequence distinguishable from endogenous nucleic acids found in natural systems. DNA sequences encoding polypeptides can be assembled from cDNA fragments or from a series of synthetic oligonucleotides, to provide a synthetic nucleic acid which is capable of being expressed from a recombinant transcriptional unit contained in a cell or in a cell-free transcription and translation system.

[0062] The terms “recombinant expression vector,” or “DNA construct” are used interchangeably herein to refer to a DNA molecule comprising a vector and at least one insert. Recombinant expression vectors are usually generated for the purpose of expressing and / or propagating the insert(s), or for the construction of other recombinant nucleotide sequences. The insert(s) may or may not be operably linked to a promoter sequence and may or may not be operably linked to DNA regulatory sequences.

[0063] As used herein, the term "affinity" refers to the equilibrium constant for the reversible binding of two agents (e.g., an antibody and an antigen) and is expressed as a dissociation constant (KD). As used herein, the term “avidity” refers to the resistance of a complex of two or more agents to dissociation after dilution. The terms “immunoreactive” and “preferentially binds” are used interchangeably herein with respect to antibodies and / or antigen-binding fragments.

[0064] The term “binding,” as used herein (e.g., with reference to binding of a TMMP to a polypeptide (e.g., a T-cell receptor) on a T cell), refers to a non-covalent interaction between two molecules. Non- covalent binding refers to a direct association between two molecules, due to, for example, electrostatic, hydrophobic, ionic, and / or hydrogen-bond interactions, including interactions such as salt bridges and water bridges. “Affinity” refers to the strength of non-covalent binding, increased binding affinity being correlated with a lower KD. “Specific binding” generally refers to binding of a ligand to a moiety that is than its designated binding site or receptor. “Non-specific binding” generally refers to binding of a ligand to a moiety other than its designated binding site or receptor. “Covalent binding” or “covalent bond,” as used herein, refers to the formation of one or more covalent chemical binds between two different molecules.

[0065] The terms “treatment”, “treating” and the like are used herein to generally mean obtaining a desired pharmacologic and / or physiologic effect. The effect may be prophylactic in terms of completely or partially preventing a disease or symptom thereof and / or may be therapeutic in terms of a partial or complete cure for a disease and / or adverse effect attributable to the disease. “Treatment” as used herein covers any treatment of a disease or symptom in a mammal, and includes: (a) preventing the disease or symptom from occurring in a subject which may or may not be predisposed to acquiring the disease or symptom but has not yet been diagnosed as having it; (b) inhibiting the disease or one or more symptoms associated with the disease, e.g., arresting its development; and / or (c) relieving the disease, i.e., causing regression of the disease. The therapeutic agent may be administered before, during and / or after the onset of disease or injury. The treatment of ongoing disease, where the treatment stabilizes or reduces the undesirable clinical symptoms of the patient, is of particular interest. Such treatment is desirably performed prior to complete loss of function in the affected tissues. The subject therapy will desirably be administered during the symptomatic stage of the disease, and in some cases after the symptomatic stage of the disease.

[0066] The terms “individual,” “subject,” “host,” and “patient,” are used interchangeably herein and refer to any mammalian subject for whom diagnosis, treatment, or therapy is desired. Mammals include, e.g., humans, non-human primates, rodents (e.g., rats; mice), lagomorphs (e.g., rabbits), ungulates (e.g., cows, sheep, pigs, horses, goats, and the like), etc. Unless otherwise indicated, the terms “individual,” “subject,” “host,” and “patient,” refer to a human.

[0067] The terms "antibodies" and “immunoglobulin” include antibodies or immunoglobulins of any isotype, fragments of antibodies that retain specific binding to antigen, including, but not limited to, Fab, Fv, scFv, and Fd fragments, chimeric antibodies, humanized antibodies, single-chain antibodies (scAb), single domain antibodies (dAb), single domain heavy chain antibodies, a single domain light chain antibodies, nanobodies, bi-specific antibodies, multi-specific antibodies, and fusion proteins comprising an antigen-binding (also referred to herein as antigen binding) portion of an antibody and a non-antibody protein. The antibodies can be detectably labeled, e.g., with a radioisotope, an enzyme that generates a detectable product, a fluorescent protein, and the like. The antibodies can be further conjugated to other moieties, such as members of specific binding pairs, e.g., biotin (member of biotin-avidin specific binding pair), and the like. Also encompassed by the term are Fab’, Fv, F(ab’)2, and or other antibody fragments that retain specific binding to antigen, and monoclonal antibodies. As used herein, a monoclonal antibody is an antibody produced by a group of identical cells, all of which were produced from a single cell by repetitive cellular replication. That is, the clone of cells only produces a single antibody species. While a monoclonal antibody can be produced using hybridoma production technology, other production methods known to those skilled in the art can also be used (e.g., antibodies derived from antibody phage display libraries). An antibody can be monovalent or bivalent. An antibodycan be an Ig monomer, which is a “Y-shaped” molecule that consists of four polypeptide chains: two heavy chains and two light chains connected by disulfide bonds.

[0068] The term "nanobody" (Nb), as used herein, refers to the smallest antigen binding fragment or single variable domain (VHH) derived from naturally occurring heavy chain antibody and is known to the person skilled in the art. They are derived from heavy chain only antibodies, seen in camelids (Hamers- Casterman et al. (1993) Nature 363:446; Desmyter et al. (1996) Nature Structural Biol.3:803; and Desmyter et al. (2015) Curr. Opin. Struct. Biol. 32:1). In the family of "camelids" immunoglobulins devoid of light polypeptide chains are found. "Camelids" comprise old world camelids (Camelus bactrianus and Camelus dromedarius) and new world camelids (for example, Llama paccos, Llama glama, Llama guanicoe and Llama vicugna). A single variable domain heavy chain antibody is referred to herein as a nanobody or a VHH antibody.

[0069] "Antibody fragments" comprise a portion of an intact antibody, for example, the antigen binding or variable region of the intact antibody. Examples of antibody fragments include Fab, Fab', F(ab')2, and Fv fragments; diabodies; linear antibodies (Zapata et al., Protein Eng.8(10): 1057-1062 (1995)); domain antibodies (dAb; Holt et al. (2003) Trends Biotechnol. 21:484); single-chain antibody molecules; and multi-specific antibodies formed from antibody fragments. Papain digestion of antibodies produces two identical antigen-binding fragments, called "Fab" fragments, each with a single antigen-binding site, and a residual "Fc" fragment, a designation reflecting the ability to crystallize readily. Pepsin treatment yields an F(ab')2fragment that has two antigen combining sites and is still capable of cross-linking antigen.

[0070] "Fv" is the minimum antibody fragment that contains a complete antigen-recognition and - binding site. This region consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. It is in this configuration that the three CDRS of each variable domain interact to define an antigen-binding site on the surface of the VH-VL dimer. Collectively, the six CDRs confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of an Fv comprising only three CDRs specific for an antigen) has the ability to recognize and bind antigen, although at a lower affinity than the entire binding site.

[0071] The “Fab” fragment also contains the constant domain of the light chain and the first constant domain (CH1) of the heavy chain. Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxyl terminus of the heavy chain CH1 domain including one or more cysteines from the antibody hinge region. Fab'-SH is the designation herein for Fab' in which the cysteine residue(s) of the constant domains bear a free thiol group. F(ab')2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.

[0072] The "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains. Depending on the amino acid sequence of the constant domain of their heavy chains, immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM, and several of these classes can be further divided into subclasses (isotypes), e.g., IgG1, IgG2, IgG3, IgG4, IgA, and IgA2. The subclasses can be further divided into types, e.g., IgG2a and IgG2b.

[0073] "Single-chain Fv" or "sFv" or “scFv” antibody fragments comprise the VH and VL domains of antibody, wherein these domains are present in a single polypeptide chain. In some embodiments, the Fv polypeptide further comprises a polypeptide linker between the VH and VL domains, which enables the sFv to form the desired structure for antigen binding. For a review of sFv, see Pluckthun in The Pharmacology of Monoclonal Antibodies, vol.113, Rosenburg and Moore eds., Springer-Verlag, New York, pp.269-315 (1994).

[0074] The term "diabodies" refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (VH) connected to a light-chain variable domain (VL) in the same polypeptide chain (VH-VL). By using a linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites. Diabodies are described more fully in, for example, EP 404,097; WO 93 / 11161; and Hollinger et al. (1993) Proc. Natl. Acad. Sci. USA 90:6444-6448.

[0075] As used herein, the term “CDR” or “complementarity determining region” is intended to mean the non-contiguous antigen combining sites found within the variable region of both heavy and light chain polypeptides. CDRs have been described by Kabat et al (1977) J. Biol. Chem.252:6609; Kabat et al., U.S. Dept. of Health and Human Services, “Sequences of proteins of immunological interest” (1991) (also referred to herein as Kabat 1991); by Chothia et al. (1987) J. Mol. Biol.196:901 (also referred to herein as Chothia 1987); and MacCallum et al. (1996) J. Mol. Biol.262:732, where the definitions include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or grafted antibodies or variants thereof is intended to be within the scope of the term as defined and used herein. The amino acid residues, which encompass the CDRs, as defined by each of the above cited references are set forth below in Table 1 as a comparison.Table 1: CDR Definitions

[0076] As used herein, the terms “CDR-L1”, “CDR-L2”, and “CDR-L3” refer, respectively, to the first, second, and third CDRs in a light chain variable region. The terms “CDR-L1”, “CDR-L2”, and “CDR- L3” may be used interchangeably with “VL CDR1,” “VL CDR2,” and “VL CDR3,” respectively. As used herein, the terms “CDR-H1”, “CDR-H2”, and “CDR-H3” refer, respectively, to the first, second, and third CDRs in a heavy chain variable region. The terms “CDR-H1”, “CDR-H2”, and “CDR-H3” may be used interchangeably with “VH CDR1,” “VH CDR2,” and “VH CDR3,” respectively. As used herein, the terms “CDR-1”, “CDR-2”, and “CDR-3” refer, respectively, to the first, second and third CDRs of either chain’s variable region.

[0077] As used herein, the term “framework,” when used in reference to an antibody variable region, is intended to mean all amino acid residues outside the CDR regions within the variable region of an antibody. A variable region framework is generally a discontinuous amino acid sequence between about 100-120 amino acids in length but is intended to reference only those amino acids outside of the CDRs. As used herein, the term “framework region” is intended to mean each domain of the framework that is separated by the CDRs.

[0078] Unless indicated otherwise, the term “substantially” is intended to encompass both “wholly” and “largely but not wholly”. For example, an Ig Fc that “substantially does not induce ADCC” means an Ig Fc that induces no ADCC at all or that largely does not induce ADCC.

[0079] As used herein, the term “about” used in connection with an amount indicates that the amount can vary by 10% of the stated amount. For example, “about 100” means an amount of from 90-110. Where about is used in the context of a range, the “about” used in reference to the lower amount of the range means that the lower amount includes an amount that is 10% lower than the lower amount of the range, and “about” used in reference to the higher amount of the range means that the higher amount includes an amount 10% higher than the higher amount of the range. For example, from about 100 to about 1000 means that the range extends from 90 to 1100.

[0080] As used herein, the term “MHC heavy chain polypeptide” means collectively the domains of an MHC heavy chain polypeptide that are present in a TMMP. For example, an MHC heavy chain polypeptide can comprise α1, α2 and α3 domains.

[0081] Before the present disclosure is further described, it is to be understood that this disclosure is not limited to particular embodiments described, as such may, of course, vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope will be limited only by the appended claims.

[0082] Where a range of values is provided, it is understood that each intervening value, to the tenth of the unit of the lower limit unless the context clearly dictates otherwise, between the upper and lower limit of that range and any other stated or intervening value in that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges, and are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. Where the stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included in the disclosure.

[0083] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can also be used in the practice or testing of the present disclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference to disclose and describe the methods and / or materials in connection with which the publications are cited.

[0084] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. Thus, for example, reference to a “T-cell modulatory polypeptide” includes a plurality of such polypeptides and reference to “the immunomodulatory polypeptide” includes reference to one or more immunomodulatory polypeptides and equivalents thereof known to those skilled in the art, and so forth. It is further noted that the claims may be drafted to exclude any optional element. As such, this statement is intended to serve as antecedent basis for use of such exclusive terminology as “solely,” “only” and the like in connection with the recitation of claim elements, or use of a “negative” limitation.

[0085] It is appreciated that certain features of the disclosure, which are, for clarity, described in the context of separate embodiments, may also be provided in combination in a single embodiment. Conversely, various features of the disclosure, which are, for brevity, described in the context of a single embodiment, may also be provided separately or in any suitable sub-combination. All combinations of the embodiments pertaining to the disclosure are specifically embraced by the present disclosure and are disclosed herein just as if each and every combination was individually and explicitly disclosed. Inaddition, all sub-combinations of the various embodiments and elements thereof are also specifically embraced by the present disclosure and are disclosed herein just as if each and every such sub- combination was individually and explicitly disclosed herein.

[0086] The publications discussed herein are provided solely for their disclosure. Nothing herein is to be construed as an admission that the present disclosure is not entitled to antedate such publication. Further, the dates of publication provided may be different from the actual publication dates which may need to be independently confirmed. DETAILED DESCRIPTION

[0087] The present disclosure provides T-cell modulatory multimeric polypeptides (TMMPs) that comprise (i) an optional immunomodulatory polypeptide such as a variant IL-2 polypeptide, (ii) class I HLA major histocompatibility complex (MHC) polypeptides (a class I HLA heavy chain polypeptide and a β2 microglobulin polypeptide), (iii) a peptide that presents an epitope to a T-cell receptor, which together with the class I MHC polypeptides forms a peptide-MHC complex (pMHC), (iv) a tumor- targeting polypeptide, and (v) an optional Ig Fc polypeptide or other scaffold.

[0088] A TMMP is useful for modulating the activity of a T cell, and for modulating an immune response in an individual. Importantly, the TMMPs disclosed herein are useful for “redirecting” a patient’s repertoire of anti-pathogenic (e.g., antiviral) T cells to attack and kill cancer cells. The TMMPs achieve this result by (i) binding to the patient’s cancer cells via the TMMP’s tumor-targeting polypeptide (TTP) that is specific for and binds an antigen on the cancer cell, and (ii) presenting a pMHC that mimics a pMHC on the surface of a pathogen-infected cell, e.g., a cell infected by a virus, bacteria or other microorganism that can cause disease. In this way, TMMPs effectively “paint” the cancer cells to make them appear like pathogen-infected cells. The “painted” cancer cells are then susceptible to being attacked and killed by the patient’s existing repertoire of anti-pathogenic T cells that have a T cell receptor (TCR) that recognizes and binds to the pMHC presented by the TMMP.

[0089] For example, TMMPs can have a TTP that binds to an antigen on a cancer cell, and a pMHC that mimics the pMHC of a cell that has been infected with a virus such as a SARS-CoV-2 virus. The patient’s existing repertoire of T cells against a SARS-CoV-2 virus-infected cells are thus re-directed to the patient’s cancer cells where the T cells can be activated through binding of the TCR of T cells to the pMHC of the TMMP. Activation of the T cells can lead to release of their cytotoxic components and killing of the cancer cells. In such cases, the patient’s repertoire of T cells specific the SARS-CoV-2 virus can be the result of one or more of a prior SARS-CoV-2 infection, vaccination with a vaccine that elicits T cells having a TCR that is specific for the pMHC of the TMMP, and / or prior treatment with a T cell modulatory protein designed to prime and / or increase the patient’s T cells that have a TCR that specifically binds the pMHC of the TMMP.

[0090] The TMMPs disclosed herein are useful for any patient having a hematological cancer or solid tumor, including patients whose cancers have evaded their immune system through HLA loss, which prevents the patient’s own cancer-specific T cells from recognizing and killing the cancer cells. T-CELL MODULATORY MULTIMERIC POLYPEPTIDES

[0091] The present disclosure provides a T-cell modulatory multimeric polypeptide (TMMP) comprising: a) a first polypeptide; and b) a second polypeptide, wherein the TMMP comprises: a peptide epitope (defined below); a first major histocompatibility complex (MHC) polypeptide; a second MHC polypeptide; one or more immunomodulatory polypeptides; an immunoglobulin (Ig) Fc polypeptide or a non-Ig scaffold; and a tumor targeting polypeptide (TTP).

[0092] In some cases, a TMMP comprises a heterodimer (comprises two different separate polypeptide chains) comprising: a) a first polypeptide; and b) a second polypeptide, wherein the TMMP comprises: a peptide epitope; a first MHC polypeptide; a second MHC polypeptide; one or more immunomodulatory polypeptides; an Ig Fc polypeptide or a non-Ig scaffold; and a TTP. A TMMP can comprise a single heterodimer. A TMMP can comprise two copies of a heterodimer; i.e., a TMMP can be a homodimer of a heterodimer. The two copies of a heterodimer can be linked to one another, e.g., by disulfide bonds (e.g., disulfide bonds between Ig Fc polypeptides present in the heterodimers)

[0093] In some cases, a TMMP comprises one or more heterodimers, each heterodimer comprising: a) a first polypeptide comprising a first MHC polypeptide; and b) a second polypeptide comprising a second MHC polypeptide, wherein the first polypeptide or the second polypeptide comprises a peptide epitope (defined below), wherein the first polypeptide and / or the second polypeptide comprises one or more immunomodulatory polypeptides that can be the same or different; and an Ig Fc polypeptide or a non-Ig scaffold. The first or the second polypeptide also includes a TTP. In some cases, the TTP is at the C- terminus of the Ig Fc polypeptide or the non-Ig scaffold. In some cases, the TTP is at the N-terminus of the polypeptide that comprises the Ig Fc polypeptide. In some cases, the TTP is at the C-terminus of the polypeptide that comprises the peptide epitope.

[0094] The peptide epitope present in a TMMP presents a SARS-CoV-2 peptide (e.g., a SARS-CoV-2 encoded peptide). As used herein, the term “peptide epitope” means a peptide that, when complexed with MHC polypeptides, presents an epitope to a T-cell receptor (TCR). When complexed with MHC polypeptides, a peptide epitope can present one or more epitopes to one or more TCRs.

[0095] In some cases, a TMMP includes: i) a SARS-CoV-2 peptide; and iii) a TTP that targets a cancer- associated antigen. Such a TMMP binds a cancer cell that expresses the cancer-associated antigen targeted by the TTP. The TMMP modulates the activity of a T-cell specific for the virus epitope present in the TMMP. For example, in some cases, the TMMP increases proliferation and / or cytotoxic activity ofa T-cell specific for the virus epitope present in the TMMP. Contacting a T-cell specific for the virus epitope present in the TMMP can increase cytotoxic activity of the T cell toward a cancer cell expressing the cancer-associated antigen that is targeted by the TTP present in the TMMP.

[0096] TMMPs comprising a pMHC that has a SARS-CoV-2 epitope are particularly useful because the high rate of SARS-CoV-2 infections worldwide and the availability of commercial vaccines means that most patients either already have, or can be induced to have through vaccination, a repertoire of T cells that can recognize a pMHC having a SARS-CoV-2 epitope. As noted above, however, a pMHC of a TMMP alternatively could present a peptide epitope from other virus or pathogenic microorganisms against which the patient has an existing repertoire of T cells. For example, the pMHC could present an epitope that would be recognized by T cells that result from administration of commonly available vaccines that provide immunity against pathogens such as tetanus, flu or other available vaccines. There are numerous commercially available vaccines against pathogens, e.g., Cholera, Dengue, Diphtheria, Hepatitis, Haemophilus influenzae type b (Hib), Human papillomavirus (HPV), Influenza, Japanese encephalitis, Malaria, Measles, Meningococcal meningitis, Mumps, Pertussis, Pneumococcal disease, Poliomyelitis, Rabies, Rotavirus, Rubella, Tetanus, Tick-borne encephalitis, Tuberculosis, Typhoid, Varicella and Yellow Fever, with numerous other in development, e.g., Enterotoxigenic Escherichia coli, Group B Streptococcus (GBS), Herpes Simplex Virus, HIV-1, Malaria, Neisseria gonorrhoeae, Nontyphoidal Salmonella Disease, Norovirus, Paratyphoid fever, Respiratory Syncytial Virus (RSV), Schistosomiasis Disease, Shigella, Group A Streptococcus (GAS), Tuberculosis, and other Influenza vaccines. Alternatively, (or additionally), prior to administering the TMMP, one or more doses of a T cell modulatory polypeptide can be administered to the patient, in order to prime and activate and / or induce proliferation of (and thereby increase the number of) CD8+T cells that recognize the desired pMHC in the TMMP comprising the TTP. See, e.g., the T cell modulatory polypeptides described in WO2021 / 195108, published September 30, 2021 (Cue Biopharma, Inc.), WO 2021 / 195411, published September 30, 2021 (Cue Biopharma, Inc.), and WO 2022 / 015880, published January 20, 2022 (Cue Biopharma, Inc.).

[0097] In some cases, a TMMP comprises one or more heterodimers, each heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope (a SARS-CoV-2 peptide); and ii) a first MHC polypeptide; b) a second polypeptide comprising a second MHC polypeptide; c) one or more immunomodulatory polypeptides, where the first and / or the second polypeptide comprises the at least one (i.e., one or more) immunomodulatory polypeptide; d) an Ig Fc polypeptide or a non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and e) a polypeptide that targets a cancer cell (a tumor-targeting polypeptide; “TTP”). In some cases, at least one of the one or more immunomodulatory polypeptides comprises an amino acid sequence of a wild- type immunomodulatory polypeptide. In some cases, at least one of the one or more immunomodulatorypolypeptides is a variant immunomodulatory polypeptide that binds to but exhibits reduced affinity to a cognate co-immunomodulatory polypeptide compared to the affinity of a corresponding wild-type immunomodulatory polypeptide for the cognate co-immunomodulatory polypeptide. In some cases, the peptide epitope present in a TMMP presents an infectious disease-associated epitope (e.g., a virus- encoded peptide).

[0098] A TMMP comprises a tumor targeting polypeptide, i.e., a polypeptide that targets a cancer- associated epitope displayed on the surface of a cancer cell.

[0099] In some cases, a TMMP is:

[0100] A) a heterodimer comprising: a) a first polypeptide comprising a first MHC polypeptide; and b) a second polypeptide comprising a second MHC polypeptide, wherein the first polypeptide or the second polypeptide comprises a SARS-CoV-2 peptide, wherein the first polypeptide and / or the second polypeptide comprises one or more immunomodulatory polypeptides that can be the same or different, and wherein at least one of the one or more immunomodulatory polypeptides may be a wild-type immunomodulatory polypeptide or a variant of a wild-type immunomodulatory polypeptide; wherein the first polypeptide and / or the second polypeptide comprises an Ig Fc polypeptide or a non-Ig scaffold; and wherein the first and / or the second polypeptide comprises a TTP; or

[0101] B) a heterodimer comprising: a) a first polypeptide comprising a first MHC polypeptide; and b) a second polypeptide comprising a second MHC polypeptide, wherein the first polypeptide or the second polypeptide comprises a SARS-CoV-2 peptide; wherein the first polypeptide and / or the second polypeptide comprises one or more immunomodulatory polypeptides that can be the same or different,

[0102] wherein at least one of the one or more immunomodulatory polypeptides is a variant of a wild-type immunomodulatory polypeptide,

[0103] wherein at least one of the one or more immunomodulatory polypeptides is a variant immunomodulatory polypeptide that exhibits reduced affinity to a cognate co-immunomodulatory polypeptide compared to the affinity of a corresponding wild-type immunomodulatory polypeptide for the cognate co-immunomodulatory polypeptide.; and

[0104] wherein the first polypeptide and / or the second polypeptide comprises an Ig Fc polypeptide or a non-Ig scaffold; and wherein the first and / or the second polypeptide comprises a TTP; or

[0105] C) a heterodimer comprising: a) a first polypeptide comprising, in order from N-terminus to C-terminus: i) a SARS-CoV-2 peptide; ii) a first MHC polypeptide; and b) a second polypeptide comprising, in order from N-terminus to C-terminus: i) a second MHC polypeptide; and ii) optionally an immunoglobulin (Ig) Fc polypeptide or a non-Ig scaffold, wherein the TMMP comprises one or more immunomodulatory polypeptides that can be the same or different, wherein at least one of the one ormore immunomodulatory polypeptides is: A) at the C-terminus of the first polypeptide; B) at the N- terminus of the second polypeptide; C) at the C-terminus of the second polypeptide; or D) at the C- terminus of the first polypeptide and at the N-terminus of the second polypeptide, and wherein at least one of the one or more immunomodulatory polypeptides may be a wild-type immunomodulatory polypeptide or a variant of a wild-type immunomodulatory polypeptide; and

[0106] optionally wherein at least one of the one or more immunomodulatory polypeptides is a variant immunomodulatory polypeptide that exhibits reduced affinity to a cognate co- immunomodulatory polypeptide compared to the affinity of a corresponding wild-type immunomodulatory polypeptide for the cognate co-immunomodulatory polypeptide.

[0107] In some cases, an immunomodulatory polypeptide present in a TMMP comprises a wild- type (naturally-occurring) amino acid sequence.

[0108] In some cases, an immunomodulatory polypeptide present in a TMMP binds to its cognate co-immunomodulatory polypeptide with an affinity that is less than the affinity of a corresponding wild-type immunomodulatory polypeptide for the cognate co-immunomodulatory polypeptide.

[0109] The combination of the reduced affinity of the immunomodulatory polypeptide for its cognate co-immunomodulatory polypeptide, and the affinity of the epitope for a TCR, provides for enhanced selectivity of a TMMP. For example, a TMMP binds selectively to a first T cell that displays both: i) a TCR specific for the epitope present in the TMMP; and ii) a co-immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP, compared to binding to a second T cell that displays: i) a TCR specific for an epitope other than the epitope present in the TMMP; and ii) a co-immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP.

[0110] In some cases, a TMMP, when administered to an individual in need thereof, induces both an epitope-specific T cell response and an epitope non-specific T cell response. In other words, in some cases, a TMMP, when administered to an individual in need thereof, induces an epitope-specific T cell response by modulating the activity of a first T cell that displays both: i) a TCR specific for the peptide epitope present in the TMMP; ii) a co-immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP; and induces an epitope non-specific T cell response by modulating the activity of a second T cell that displays: i) a TCR specific for an epitope other than the epitope present in the TMMP; and ii) a co-immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP. The ratio of the epitope-specific T cell response to the epitope-non-specific T cell response is at least 2:1, at least 5:1, at least 10:1, at least 15:1, at least 20:1, at least 25:1, at least 50:1, or at least 100:1. The ratio of the epitope-specific T cell responseto the epitope-non-specific T cell response is from about 2:1 to about 5:1, from about 5:1 to about 10:1, from about 10:1 to about 15:1, from about 15:1 to about 20:1, from about 20:1 to about 25:1, from about 25:1 to about 50:1, or from about 50:1 to about 100:1, or more than 100:1. “Modulating the activity” of a T cell can include one or more of: i) activating a cytotoxic (e.g., CD8+) T cell; ii) inducing cytotoxic activity of a cytotoxic (e.g., CD8+) T cell; iii) inducing production and release of a cytotoxin (e.g., a perforin; a granzyme; a granulysin) by a cytotoxic (e.g., CD8+) T cell; iv) inhibiting activity of an autoreactive T cell; and the like. As used herein, selectively modulating refers to inducing an epitope- specific T cell response that is greater than the epitope-non-specific T cell response.

[0111] The combination of the reduced affinity of the immunomodulatory polypeptide for its cognate co-immunomodulatory polypeptide, and the affinity of the epitope for a TCR, provides for enhanced selectivity of a TMMP. Thus, for example, a TMMP binds with higher avidity to a first T cell that displays both: i) a TCR specific for the epitope present in the TMMP; and ii) a co- immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP, compared to the avidity to which it binds to a second T cell that displays: i) a TCR specific for an epitope other than the epitope present in the TMMP; and ii) a co-immunomodulatory polypeptide that binds to the immunomodulatory polypeptide present in the TMMP.

[0112] Binding affinity between an immunomodulatory polypeptide and its cognate co- immunomodulatory polypeptide can be determined by bio-layer interferometry (BLI) as described in published PCT Application WO 2020 / 243315 (Cue Biopharma, Inc.). Dimerized TMMPs

[0113] A TMMP can be dimerized; i.e., the present disclosure provides a multimeric polypeptide comprising a dimer of a TMMP. Thus, the present disclosure provides a TMMP comprising: A) a first heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope; and ii) a first major histocompatibility complex (MHC) polypeptide; and b) a second polypeptide comprising: i) a second MHC polypeptide, wherein the first heterodimer comprises one or more immunomodulatory polypeptides; and B) a second heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope; and ii) a first MHC polypeptide; and b) a second polypeptide comprising: i) a second MHC polypeptide, wherein the second heterodimer comprises one or more immunomodulatory polypeptides, and wherein the first heterodimer and the second heterodimer are covalently linked to one another. In some cases, the two TMMPs are identical to one another in amino acid sequence. In some cases, the first heterodimer and the second heterodimer are covalently linked to one another via a C-terminal region of the second polypeptide of the first heterodimer and a C-terminal region of the second polypeptide of the second heterodimer. In some cases, first heterodimer and the second heterodimer are covalently linked to one another via the C-terminal amino acid of the second polypeptide of the first heterodimer and the C- terminal region of the second polypeptide of the second heterodimer; for example, in some cases, the C-terminal amino acid of the second polypeptide of the first heterodimer and the C-terminal region of the second polypeptide of the second heterodimer are linked to one another, either directly or via a linker. The linker can be a peptide linker. The peptide linker can have a length of from 1 amino acid to 200 amino acids (e.g., from 1 amino acid (aa) to 5 aa, from 5 aa to 10 aa, from 10 aa to 25 aa, from 25 aa to 50 aa, from 50 aa to 100 aa, from 100 aa to 150 aa, or from 150 aa to 200 aa). In some cases, the peptide epitope of the first heterodimer and the peptide epitope of the second heterodimer comprise the same amino acid sequence. In some cases, the first MHC polypeptide of the first and the second heterodimer is an MHC Class I β2-microglobulin, and wherein the second MHC polypeptide of the first and the second heterodimer is an MHC Class I heavy chain. In some cases, the immunomodulatory polypeptide of the first heterodimer and the immunomodulatory polypeptide of the second heterodimer comprise the same amino acid sequence. In some cases, the immunomodulatory polypeptide of the first heterodimer and the immunomodulatory polypeptide of the second heterodimer are variant immunomodulatory polypeptides that comprise from 1 to 10 amino acid substitutions compared to a corresponding parental wild-type immunomodulatory polypeptide, and wherein the from 1 to 10 amino acid substitutions result in reduced affinity binding of the variant immunomodulatory polypeptide to a cognate co-immunomodulatory polypeptide. In some cases, the immunomodulatory polypeptide of the first heterodimer and the immunomodulatory polypeptide of the second heterodimer are each independently selected from the group consisting of IL-2, 4-1BBL, PD-L1, CD80, CD86, ICOS-L, OX-40L, FasL, JAG1 (CD339), TGFβ, CD70, and ICAM. Examples, of suitable MHC polypeptides, immunomodulatory polypeptides, and peptide epitopes are described below. The first and / or the second polypeptide comprises: i) an Ig Fc polypeptide or a non-Ig scaffold; and ii) a tumor-targeting polypeptide. Multiple disulfide bonded TMMPs

[0114] In some cases, the first polypeptide and the second polypeptide of a TMMP are linked to one another by at least two disulfide bonds (i.e., two interchain disulfide bonds). Examples of such multiple disulfide-linked TMMP are depicted schematically in FIG.18A-FIG.18C. In addition, where a TMMP comprises an IgFc polypeptide, a heterodimeric TMMP can be dimerized, such that disulfide bonds link the IgFc polypeptides in the two heterodimeric TMMPs. Such an arrangement is depicted schematically in FIG.18D, where disulfide bonds are represented by dashed lines. Unless otherwise stated, the at least two disulfide bonds described in the multiple disulfide-linked TMMPs in this section are not referring to disulfide bonds linking IgFc polypeptides in dimerized TMMPs.

[0115] As noted above, in some cases, the first polypeptide and the second polypeptide of a TMMP are linked to one another by at least two disulfide bonds (i.e., two interchain disulfide bonds). For example, in some instances, the first polypeptide and the second polypeptide of a TMMP are linked to one another by 2 interchain disulfide bonds. As another example, in some instances, the first polypeptide and the second polypeptide of a TMMP are linked to one another by 3 interchain disulfidebonds. As another example, in some instances, the first polypeptide and the second polypeptide of a TMMP are linked to one another by 4 interchain disulfide bonds.

[0116] In some cases where a peptide epitope in a first polypeptide of a TMMP is linked to a β2M polypeptide by a linker comprising a Cys, at least one of the at least two disulfide bonds links a Cys in the linker to a Cys in an MHC Class I heavy chain in the second polypeptide. In some cases, where a peptide epitope in a first polypeptide of a TMMP is linked to an MHC Class I heavy chain polypeptide by a linker, at least one of the at least two disulfide bonds links a Cys in the linker to a Cys in a β2M polypeptide present in the second polypeptide.

[0117] In some cases, a T-cell modulatory multimeric polypeptide comprises at least one heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope; and ii) first MHC polypeptide; b) a second polypeptide comprising a second MHC polypeptide; c) at least one immunomodulatory polypeptide, where the first and / or the second polypeptide comprises the at least one immunomodulatory polypeptide; d) an Ig Fc polypeptide or a non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and e) a TTP, where the first and / or the second polypeptide comprises the TTP; and where the heterodimer comprises at least two disulfide bonds (e.g., two disulfide bonds) between the first polypeptide and the second polypeptide (e.g., the heterodimer comprises: i) a first disulfide bond linking the first polypeptide and the second polypeptide; and ii) a second disulfide bond linking the first polypeptide and the second polypeptide). Expressed another way, the first polypeptide comprises a first Cys residue that forms a disulfide bond (a first disulfide bond) with a first Cys residue in the second polypeptide; and the first polypeptide comprises a second Cys residue that forms a disulfide bond (a second disulfide bond) with a second Cys residue in the second polypeptide.

[0118] In some cases, a TMMP comprises: a) a first polypeptide comprising, in order from N- terminus to C-terminus: i) a peptide epitope; ii) a peptide linker; and iii) a β2M polypeptide; and b) a second polypeptide comprising an MHC Class I heavy chain polypeptide, where one or both of the first and the second polypeptides comprises at least one immunomodulatory polypeptide, where the TMMP comprises: a) a first disulfide linkage between: i) a Cys present in the linker between the peptide epitope and the β2M polypeptide; and ii) a first Cys introduced into the MHC Class I heavy chain polypeptide; and b) at least a second disulfide linkage between the first polypeptide and the second polypeptide, where the at least a second disulfide linkage is between: i) a Cys in the first polypeptide that is C- terminal to the Cys present in the linker; and ii) a Cys in the second polypeptide that is C-terminal to the first Cys introduced into the MHC Class I heavy chain polypeptide. As noted above, the TMMP also includes: i) an Ig Fc polypeptide or a non-Ig scaffold; and ii) a TTP.

[0119] A multiple disulfide-linked TMMP (e.g., a double disulfide-linked TMMP) can comprise, for example: a) a first polypeptide comprising: i) a peptide epitope; and ii) a first MHC polypeptide,where the first polypeptide comprises a peptide linker between the peptide and the first MHC polypeptide, where the peptide linker comprises a Cys residue, and where the first MHC polypeptide is a β2M polypeptide that comprises an amino acid substitution that introduces a Cys residue; b) and a second polypeptide comprising a second MHC polypeptide, where the second MHC polypeptide is a Class I heavy chain comprising a Y84C substitution and an A236C substitution, based on the amino acid numbering of HLA-A*0201 (depicted in FIG.7A), or at corresponding positions in another Class I heavy chain allele, where the TMMP comprises a disulfide bond between the Cys residue in the peptide linker and the Cys residue at amino acid position 84 of the Class I heavy chain or corresponding position of another Class I heavy chain allele, and where the TMMP comprises a disulfide bond between the introduced Cys residue in the β2M polypeptide and the Cys at amino acid position 236 of the Class I heavy chain or corresponding position of another Class I heavy chain allele; and c) at least one immunomodulatory polypeptide, where the first and / or the second polypeptide comprises the at least one immunomodulatory polypeptide. As noted above, the TMMP also includes: i) an Ig Fc polypeptide or a non-Ig scaffold; and ii) a TTP. SARS-COV-2 PEPTIDES

[0120] As discussed above, a TMMP comprises a Betacoronavirus (e.g., SARS-CoV-2) peptide that is typically at least about 4 amino acids in length, and presents a SARS-CoV-2 epitope to a T cell when in an MHC / peptide complex (e.g., an HLA / peptide complex).

[0121] A SARS-CoV-2 peptide present in a TMMP can have a length of at least 4 amino acids, e.g., from 4 amino acids to about 25 amino acids in length (e.g., 4 amino acids (aa), 5 aa, 6 aa, 7 aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa, including within a range of from 4 to 20 amino acids, from 6 to 18 amino acids, from 8 to 15 amino acids, from 8 to 12 amino acids, from 9-11 amino acids, from 9-10 amino acids, from 5 to 10 amino acids, from 10-15 amino acids, from 10 to 20 amino acids, and from 15 to 25 amino acids in length), for example, lengths of 9, 10, 11, 12, 13 or 14 amino acids.

[0122] A SARS-CoV-2 epitope present in a TMMP is a peptide specifically bound by a T-cell, i.e., the epitope is specifically bound by an epitope-specific T cell. An epitope-specific T cell binds an epitope having a reference amino acid sequence, but does not substantially bind an epitope that differs from the reference amino acid sequence. For example, an epitope-specific T cell binds an epitope having a reference amino acid sequence, and binds an epitope that differs from the reference amino acid sequence, if at all, with an affinity that is less than 10-6M, less than 10-5M, or less than 10-4M. An epitope-specific T cell can bind an epitope for which it is specific with an affinity of at least 10-7M, at least 10-8M, at least 10-9M, or at least 10-10M.

[0123] The peptide epitope present in a TMMP is a peptide of a Betacoronavirus-encoded polypeptide. In some cases, the peptide epitope is a SARS-CoV-2 peptide (i.e., a peptide of a SARS-CoV-2-encoded polypeptide). In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2-encoded surface glycoprotein. In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2-encoded membrane glycoprotein. In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2-encoded nucleocapsid phosphoprotein.

[0124] In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of a polypeptide comprising an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to any one of the SARS-CoV-2 polypeptides depicted in FIG.13A-13J. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of one of the polypeptides depicted in FIG.13A-13J.

[0125] In some cases, a peptide present in a TMMP is any one of the peptides depicted in FIG. 14. In some cases, a peptide present in a TMMP is any one of the peptides depicted in FIG.15.

[0126] In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2- encoded surface glycoprotein. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of a polypeptide comprising an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the SARS-CoV-2 surface glycoprotein depicted in FIG.13J. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of the SARS-CoV-2 surface glycoprotein depicted in FIG.13J.

[0127] In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2- encoded surface glycoprotein, where the SARS-CoV-2 is the omicron variant (also known as the B1.1.529 variant). In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of a polypeptide comprising an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the SARS-CoV-2 surface glycoprotein depicted in FIG.13K. As shown in FIG.13K, the spike glycoprotein of the omicron variant includes the following changes: A67V, del69-70, T95I, del142-144, Y145D, del211, L212I, ins214EPE, G339D, S371L, S373P, S375F, K417N, N440K, G446S, S477N, T478K,E484A, Q493R, G496S, Q498R, N501Y, Y505H, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, L981F, where the substitutions in bold are in the receptor binding domain (RBD), where “ins” refers to an insertion, and where “del” refers to a deletion. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of the SARS-CoV-2 surface glycoprotein depicted in FIG.13K.

[0128] In some cases, a peptide present in a TMMP is a SARS-CoV-2 peptide from a SARS- CoV-2-encoded surface glycoprotein and is selected from the group consisting of: NLTTRTQL (SEQ ID NO:533), LPPAYTNSF (SEQ ID NO:590), KVFRSSVLH (SEQ ID NO:288), LPFFSNVTW (SEQ ID NO:591), PFFSNVTWF (SEQ ID NO:388), RFDNPVLPF (SEQ ID NO:389), LPFNDGVYF (SEQ ID NO:475), GVYFASTEK (SEQ ID NO:289), TEKSNIIRGW (SEQ ID NO:711), TLDSKTQSL (SEQ ID NO:534), GVYYHKNNK (SEQ ID NO:290), YYHKNNKSW (SEQ ID NO:412), VYSSANNCTF (SEQ ID NO:390), FEYVSQPFL (SEQ ID NO:661), EYVSQPFLM (SEQ ID NO:391), FVFKNIDGY (SEQ ID NO:592), TPINLVRDL (SEQ ID NO:476), LPQGFSAL (SEQ ID NO:477), LPIGINITRF (SEQ ID NO:593), INITRFQTL (SEQ ID NO:535), LLALHRSYL (SEQ ID NO:536), WTAGAAAYY (SEQ ID NO:145), YYVGYLQPRTF (SEQ ID NO:392), YLQPRTFLL (SEQ ID NO:218), YLQPRTFL (SEQ ID NO:537), SETKCTLKSF (SEQ ID NO:712), TLKSFTVEK (SEQ ID NO:291), QPTESIVRF (SEQ ID NO:594), RFPNITNLCPF (SEQ ID NO:413), GEVFNATRF (SEQ ID NO:662), NATRFASVY (SEQ ID NO:595), LYNSASFSTF (SEQ ID NO:393), NSASFSTFK (SEQ ID NO:329), RQIAPGQTGK (SEQ ID NO:292), KIADYNYKL (SEQ ID NO:219), NYNYLYRLF (SEQ ID NO:394), RLFRKSNLK (SEQ ID NO:293), KPFERDISTEI (SEQ ID NO:478), YFPLQSYGF (SEQ ID NO:395), QPYRVVVL (SEQ ID NO:479), PYRVVVLSF (SEQ ID NO:396), GPKKSTNLV (SEQ ID NO:480), TSNQVAVLY (SEQ ID NO:146), VYSTGSNVF (SEQ ID NO:397), AEHVNNSY (SEQ ID NO:721), IPIGAGICASY (SEQ ID NO:596), SPRRARSVA (SEQ ID NO:481), VASQSIIAY (SEQ ID NO:597), SIIAYTMSL (SEQ ID NO:220), LGAENSVAY (SEQ ID NO:598), AYSNNSIAIPTNF (SEQ ID NO:414), IPTNFTISV (SEQ ID NO:482), TEILPVSMTK (SEQ ID NO:330), QEVFAQVKQIY (SEQ ID NO:713), KQIYKTPPIK (SEQ ID NO:294), IYKTPPIKDF (SEQ ID NO:398), LLFNKVTLA (SEQ ID NO:221), TLADAGFIK (SEQ ID NO:295), LADAGFIKQY (SEQ ID NO:147), ADAGFIKQY (SEQ ID NO:714), VLPPLLTDEMIAQY (SEQ ID NO:148), IPFAMQMAY (SEQ ID NO:599), SSTASALGK (SEQ ID NO:331), VLNDILSRL (SEQ ID NO:222), RLDKVEAEV (SEQ ID NO:223), VEAEVQIDRL (SEQ ID NO:663), AEVQIDRLI (SEQ ID NO:664), LITGRLQSL (SEQ ID NO:538), RLQSLQTYV (SEQ ID NO:224), AEIRASANL (SEQ ID NO:665), ASANLAATK (SEQ ID NO:296), HLMSFPQSA (SEQ ID NO:225), FPQSAPHGVVF (SEQ ID NO:600), APHGVVFL (SEQ ID NO:483), VTYVPAQEK (SEQ ID NO:297), TYVPAQEKNF (SEQ ID NO:399), REGVFVSNGTHW (SEQ ID NO:715), GTHWFVTQR (SEQ ID NO:332), TVYDPLQPELDSFK (SEQID NO:333), KEIDRLNEV (SEQ ID NO:666), QELGKYEQYIKW (SEQ ID NO:716), YEQYIKWPW (SEQ ID NO:717), QYIKWPWYI (SEQ ID NO:400), FIAGLIAIV (SEQ ID NO:226), and SEPVLKGVKL (SEQ ID NO:484).

[0129] In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2- encoded membrane glycoprotein. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of a polypeptide comprising an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the SARS-CoV-2 membrane glycoprotein depicted in FIG.13D. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of the SARS-CoV-2 membrane glycoprotein depicted in FIG.13D.

[0130] In some cases, a peptide present in a TMMP is a SARS-CoV-2 peptide from a SARS- CoV-2-encoded membrane glycoprotein and is selected from the group consisting of: GTITVEELK (SEQ ID NO:302), EELKKLLEQW (SEQ ID NO:671), KLLEQWNLV (SEQ ID NO:155), FAYANRNRF (SEQ ID NO:544), YANRNRFLY (SEQ ID NO:545), SYFIASFRLF (SEQ ID NO:338), RLFARTRSM (SEQ ID NO:491), VPLHGTIL (SEQ ID NO:427), SELVIGAVIL (SEQ ID NO:602), HLRIAGHHL (SEQ ID NO:492), RIAGHHLGR (SEQ ID NO:233), KEITVATSRTL (SEQ ID NO:603), ATSRTLSYYK (SEQ ID NO:303), ASQRVAGDSGFAAY (SEQ ID NO:101), and VAGDSGFAAY (SEQ ID NO:102).

[0131] In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2- encoded nucleocapsid phosphoprotein. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of a polypeptide comprising an amino acid sequence having at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the SARS-CoV-2 nucleocapsid phosphoprotein depicted in FIG.13F. In some cases, a peptide present in a TMMP is a peptide of from 4 amino acids (aa) to 25 aa in length (e.g., 4 aa, 5 aa, 6 aa, 7, aa, 8 aa, 9 aa, 10 aa, 11 aa, 12 aa, 13 aa, 14 aa, 15 aa, 16 aa, 17 aa, 18 aa, 19 aa, 20 aa, 21 aa, 22 aa, 23 aa, 24 aa, or 25 aa in length) of the SARS-CoV-2 nucleocapsid phosphoprotein depicted in FIG.13F.

[0132] In some cases, the peptide epitope is a SARS-CoV-2 peptide from a SARS-CoV-2- encoded nucleocapsid phosphoprotein and is selected from the group consisting of: LPNNTASWF (SEQ ID NO:567), KFPRGQGVPI (SEQ ID NO:454), NTNSSPDDQIGYY (SEQ ID NO:119), SPRWYFYYL (SEQ ID NO:455), LLLDRLNQL (SEQ ID NO:192), KAYNVTQAF (SEQ ID NO:568),QELIRQGTDYKHW (SEQ ID NO:690), ASAFFGMSR (SEQ ID NO:315), SRIGMEVTPSGTW (SEQ ID NO:691), GMEVTPSGTWL (SEQ ID NO:692), TPSGTWLTY (SEQ ID NO:569), AYKTFPPTEPK (SEQ ID NO:316), and LPAADLDDF (SEQ ID NO:570).

[0133] In some cases, the peptide epitope is a SARS-CoV-2 peptide depicted in FIG. 19. In some cases, the peptide is an HLA-A*01:01-restricted peptide depicted in FIG.19. In some cases, the peptide is an HLA-A*02:01-restricted peptide depicted in FIG.19. In some cases, the peptide is an HLA-A*11:01-restricted peptide depicted in FIG.19. In some cases, the peptide is an HLA-A*24:01- restricted peptide depicted in FIG.19. In some cases, the peptide is an HLA-B*07:02-restricted peptide depicted in FIG.19. The peptides in FIG.19 may be paired with an HLA allele as shown in FIG.19 or a different HLA allele.

[0134] In some cases, the peptide epitope is RLQSLQTYV (SEQ ID NO:224). In some cases, the peptide epitope is YLQPRTFLL (SEQ ID NO:218).

[0135] In some cases, the peptide epitope is one that can be presented in a complex with a β2M polypeptide and an HLA-E polypeptide. As one non-limiting example, in some cases, the peptide epitope is a peptide of a SARS-CoV-2 Nsp13 (helicase) polypeptide (see, e.g., FIG.13C). In some cases, the peptide epitope is VMPLSAPTL (SEQ ID NO:914). In some cases, a TMMP comprises an HLA-E heavy chain polypeptide and comprises the peptide epitope VMPLSAPTL (SEQ ID NO:914). MHC POLYPEPTIDES

[0136] As noted above, a TMMP includes MHC polypeptides. For the purposes of the instant disclosure, the term “major histocompatibility complex (MHC) polypeptides” is meant to include MHC polypeptides of various species, including human MHC (also referred to as human leukocyte antigen (HLA)) polypeptides, rodent (e.g., mouse, rat, etc.) MHC polypeptides, and MHC polypeptides of other mammalian species (e.g., lagomorphs, non-human primates, canines, felines, ungulates (e.g., equines, bovines, ovines, caprines, etc.), and the like. The term “MHC polypeptide” is meant to include Class I MHC polypeptides (e.g., β-2 microglobulin and MHC class I heavy chain).

[0137] In some cases, the first MHC polypeptide is an MHC class I beta-2 microglobulin (β2M) polypeptide, and the second MHC polypeptide is an MHC class I heavy chain (H chain) (“MHC-H”)). In other instances, the first MHC polypeptide is an MHC class I heavy chain polypeptide; and the second MHC polypeptide is a β2M polypeptide. In some cases, both the β2M and MHC-H chain are of human origin; i.e., the MHC-H chain is an HLA heavy chain, or a variant thereof. Unless expressly stated otherwise, a TMMP does not include membrane anchoring domains (transmembrane regions) of an MHC class I heavy chain, or a part of MHC class I heavy chain sufficient to anchor the resulting TMMP to a cell (e.g., eukaryotic cell such as a mammalian cell) in which it is expressed. In some cases, the MHC class I heavy chain present in a TMMP does not include a signal peptide, a transmembranedomain, or an intracellular domain (cytoplasmic tail) associated with a native MHC class I heavy chain. Thus, e.g., in some cases, the MHC class I heavy chain present in a TMMP includes only the α1, α2, and α3 domains of an MHC class I heavy chain polypeptide. In some cases, the MHC class I heavy chain present in a TMMP has a length of from about 270 amino acids (aa) to about 290 aa. In some cases, the MHC class I heavy chain present in a TMMP has a length of 270 aa, 271 aa, 272 aa, 273 aa, 274 aa, 275 aa, 276 aa, 277 aa, 278 aa, 279 aa, 280 aa, 281 aa, 282 aa, 283 aa, 284 aa, 285 aa, 286 aa, 287 aa, 288 aa, 289 aa, or 290 aa. MHC class I heavy chains

[0138] In some cases, an MHC class I heavy chain polypeptide present in a TMMP comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to all or part (e.g., 50, 75, 100, 150, 200, or 250 contiguous amino acids) of the amino acid sequence of any of the human HLA heavy chain polypeptides depicted in FIGs.3-11. In some cases, the MHC class I heavy chain has a length of 270 aa, 271 aa, 272 aa, 273 aa, 274 aa, 275 aa, 276 aa, 277 aa, 278 aa, 279 aa, 280 aa, 281 aa, 282 aa, 283 aa, 284 aa, 285 aa, 286 aa, 287 aa, 288 aa, 289 aa, or 290 aa. In some cases, the MHC class I heavy chain has a length of from 270 aa to 280 aa. In some cases, an MHC class I heavy chain polypeptide present in a TMMP comprises 1-30, 1-5, 5-10, 10-15, 15-20, 20-25 or 25-30 amino acid insertions, deletions, and / or substitutions (in addition to those locations indicated as being variable in the heavy chain consensus sequences) of any one of the amino acid sequences depicted in FIG.3-11. As noted above, the MHC class I heavy chain typically does not include transmembrane or cytoplasmic domains.

[0139] In some cases, an MHC polypeptide of a TMMP is a human MHC polypeptide, where human MHC polypeptides are also referred to as “human leukocyte antigen” (“HLA”) polypeptides. In some cases, an MHC polypeptide of a TMMP is a Class I HLA polypeptide, e.g., a β2-microglobulin polypeptide, or a Class I HLA heavy chain polypeptide. Class I HLA heavy chain polypeptides include HLA-A heavy chain polypeptides, HLA-B heavy chain polypeptides, HLA-C heavy chain polypeptides, HLA-E heavy chain polypeptides, HLA-F heavy chain polypeptides, and HLA-G heavy chain polypeptides.

[0140] In some cases, a TMMP comprises an HLA-A heavy chain polypeptide. The HLA-A heavy chain peptide sequences, or portions thereof, that may be that may be incorporated into a TMMP include, but are not limited to, the alleles: A*0101, A*0201, A*0301, A*1101, A*2301, A*2402, A*2407, A*3303, and A*3401. Any of those alleles may comprise a mutation at one or more of positions 84, 139, and 236 (as shown in FIG. 3-6 selected from: a tyrosine to alanine at position 84 (Y84A); a tyrosine to cysteine at position 84 (Y84C); an alanine to cysteine at position 139 (A139C); and an alanine to cysteine substitution at position 236 (A236C). In addition, HLA-A sequence having at least 75% (e.g., atleast 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%) or 100% amino acid sequence identity to all or part (e.g., 50, 75, 100, 150, 200, or 250 contiguous amino acids) of the sequence of those HLA-A alleles may also be employed (e.g., it may comprise 1-25, 1-5, 5-10, 10-15, 15-20, 20-25, or 25-30 amino acid insertions, deletions, and / or substitutions). Some examples are provided below. HLA-A02

[0141] In some cases, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.3A. In some cases, an HLA-A heavy chain polypeptide suitable for inclusion in a TMMP comprises the amino acid sequence depicted in FIG.3A. This HLA-A heavy chain polypeptide is also referred to as “HLA- A*0201” or simply “HLA-A02.” As described in WO 2018 / 119114 and WO 2020 / 132138, TMMPs can comprise one or more mutations from the wild-type HLA-A02, including to provide Cys residues that can form disulfide bonds, e.g., (i) between the β2M and MHC class I heavy chain, and / or (ii) between the MHC heavy chain and a linker that joins the peptide epitope to the β2M polypeptide, and / or (iii) an intrachain disulfide bond within the MHC class I heavy chain polypeptide.

[0142] To facilitate the formation of such disulfide bonds, one or more non-naturally occurring Cys residues can be provided in the heavy chain polypeptide. For example, the MHC class I heavy chain polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to A) the HLA-A02 (Y84C; A236C) amino acid sequence depicted in FIG.3C, where amino acid 84 is a Cys, and where amino acid 236 is a Cys. In some cases, the Cys-236 forms an interchain disulfide bond with Cys-12 of a variant β2M polypeptide that comprises an R12C substitution; or B) the HLA-A02 (Y84A; A236C) amino acid sequence: depicted in FIG.3D, where amino acid 84 is Ala and amino acid 236 is Cys. In some cases, the Cys-236 forms an interchain disulfide bond with Cys-12 of a variant β2M polypeptide that comprises an R12C substitution; or C) the HLA-A02 (Y84C; A139C) amino acid sequence depicted in FIG.3E, where amino acid 84 is Cys and amino acid 139 is Cys.

[0143] As noted above, in some cases the MHC class I heavy chain polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the HLA-A02 (Y84A; A236 wild-type) amino acid sequence depicted in FIG.3B, where amino acid 84 is a Tyr, and amino acid 236 is an Ala.HLA-A11 (HLA-A*1101)

[0144] In some cases, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.4A. In some cases, an HLA-A heavy chain polypeptide suitable for inclusion in a TMMP comprises the amino acid sequence depicted in FIG.4A. This HLA-A heavy chain polypeptide is also referred to as “HLA- A*1101” or simply “HLA-A11.” Variants can include, e.g., the MHC class I heavy chain polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to: A) the HLA-A11 (Y84A; A236 wild-type) amino acid sequence depicted in FIG. 4B, where amino acid 84 is a Tyr, and amino acid 236 is an Ala; B) the HLA-A11 (Y84C; A236C) amino acid sequence depicted in FIG.4C, where amino acid 84 is a Cys, and where amino acid 236 is a Cys; C) the HLA-A11 (Y84A; A236C) amino acid sequence: depicted in FIG.4D, where amino acid 84 is Ala and amino acid 236 is Cys. In some cases, the Cys-236 forms an interchain disulfide bond with Cys-12 of a variant β2M polypeptide that comprises an R12C substitution; and D) the HLA-A11 (Y84C; A139C) amino acid sequence depicted in FIG.4E, where amino acid 84 is Cys and amino acid 139 is Cys. In some cases, Cys-84 forms an intrachain disulfide bond with Cys-139. HLA-A24 (HLA-A*2402)

[0145] In some cases, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.5A. In some cases, an HLA-A heavy chain polypeptide suitable for inclusion in a TMMP comprises the amino acid sequence depicted in FIG.5A. This HLA-A heavy chain polypeptide is also referred to as “HLA- A*2402” or simply “HLA-A24.” Variants can include, e.g., the MHC class I heavy chain polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to: A) the HLA-A24 (Y84A; A236 wild-type) amino acid sequence depicted in FIG. 5B, where amino acid 84 is a Tyr, and amino acid 236 is an Ala; B) the HLA-A24 (Y84C; A236C) amino acid sequence depicted in FIG.5C, where amino acid 84 is a Cys, and where amino acid 236 is a Cys;C) the HLA-A24 (Y84A; A236C) amino acid sequence: depicted in FIG.5D, where amino acid 84 is Ala and amino acid 236 is Cys. In some cases, the Cys-236 forms an interchain disulfide bond with Cys-12 of a variant β2M polypeptide that comprises an R12C substitution; and D) the HLA-A24 (Y84C; A139C) amino acid sequence depicted in FIG.5E, where amino acid 84 is Cys and amino acid 139 is Cys. In some cases, Cys-84 forms an intrachain disulfide bond with Cys-139. HLA-A33 (HLA-A*3303)

[0146] In some cases, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.6A. In some cases, an HLA-A heavy chain polypeptide suitable for inclusion in a TMMP comprises the amino acid sequence depicted in FIG.6A. This HLA-A heavy chain polypeptide is also referred to as “HLA- A*3303” or simply “HLA-A33.” Variants can include, e.g., the MHC class I heavy chain polypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to: A) the HLA-A24 (Y84A; A236 wild-type) amino acid sequence depicted in FIG. 6B, where amino acid 84 is a Tyr, and amino acid 236 is an Ala; B) the HLA-A24 (Y84C; A236C) amino acid sequence depicted in FIG.6C, where amino acid 84 is a Cys, and where amino acid 236 is a Cys; C) the HLA-A24 (Y84A; A236C) amino acid sequence: depicted in FIG.6D, where amino acid 84 is Ala and amino acid 236 is Cys. In some cases, the Cys-236 forms an interchain disulfide bond with Cys-12 of a variant β2M polypeptide that comprises an R12C substitution; and D) the HLA-A24 (Y84C; A139C) amino acid sequence depicted in FIG.6E, where amino acid 84 is Cys and amino acid 139 is Cys. In some cases, Cys-84 forms an intrachain disulfide bond with Cys-139.

[0147] FIGs.7-9 provide alignments of mature HLA class I heavy chain amino acid sequences (without leader sequences or transmembrane domains or intracellular domains). The aligned amino acid sequences in FIG.7A are HLA-A class I heavy chains of the following alleles: A*0101, A*0201, A*0301, A*1101, A*2301, A*2402, A*2407, A*3303, and A*3401. The aligned amino acid sequences in FIG. 8A are HLA-B class I heavy chains of the following alleles: B*0702, B*0801, B*1502, B*3802, B*4001, B*4601, and B*5301. The aligned amino acid sequences in FIG. 9A are HLA-C class I heavy chains of the following alleles: C*0102, C*0303, C*0304, C*0401, C*0602, C*0701, C*0801, and C*1502. Indicated in the alignments are the locations (84 and 139 of the mature proteins) where cysteine residues may be introduced (e.g., by substitution) for the formation of a disulfide bond to stabilize theHLA H chain – β2M complex. Also shown in the alignment is position 236 (of the mature polypeptide), which may be substituted by a cysteine residue that can form an inter-chain disulfide bond with β2M (e.g., at aa 12). The boxes flanking residues 84, 139 and 236 show the groups of five amino acids on either sides of those six sets of five residues, denoted aac1 (for “amino acid cluster 1”), aac2 (for “amino acid cluster 2”), aac3 (for “amino acid cluster 3”), aac4 (for “amino acid cluster 4”), aac5 (for “amino acid cluster 5”), and aac6 (for “amino acid cluster 6”), that may be replaced by 1 to 5 amino acids selected independently from (i) any naturally occurring amino acid or (ii) any naturally occurring amino acid except proline or glycine.

[0148] FIGs.7A, 8A, and 9A provide alignments of the amino acid sequences of mature HLA- A, -B, and -C class I heavy chains, respectively. The sequences are provided for the extracellular portion of the mature protein (without leader sequences or transmembrane domains or intracellular domains). The positions of aa residues 84, 139, and 236 and their flanking residues (aac1 to aac6) that may be replaced by 1 to 5 amino acids selected independently from (i) any naturally occurring amino acid or (ii) any naturally occurring amino acid except proline or glycine ae also shown. FIG.7B, 8B, and 9B provide consensus amino acid sequences for the HLA-A, -B, and -C sequences, respectively, provide in FIG.7A, 8A, and 9A. The consensus sequences show the variable amino acid positions as “X” residues sequentially numbered and the locations of amino acids 84, 139 and 236 double underlined.

[0149] With regard to FIG.7A, in some cases: i) aac1 (amino acid cluster 1) may be the amino acid sequence GTLRG (SEQ ID NO:915) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., L replaced by I, V, A or F); ii) aac2 (amino acid cluster 2) may be the amino acid sequence YNQSE (SEQ ID NO:916) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., N replaced by Q, Q replaced by N, and / or E replaced by D); iii) aac3 (amino acid cluster 3) may be the amino acid sequence TAADM (SEQ ID NO:917) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., T replaced by S, A replaced by G, D replaced by E, and / or M replaced by L, V, or I); iv) aac4 (amino acid cluster 4) may be the amino acid sequence AQTTK (SEQ ID NO:918) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., A replaced by G, Q replaced by N, or T replaced by S, and or K replaced by R or Q); v) aac5 (amino acid cluster 5) may be the amino acid sequence VETRP (SEQ ID NO:919) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., V replaced by I or L, E replaced by D, T replaced by S, and / or R replaced by K); and / or vi) aac6 (amino acid cluster 6) may be the amino acid sequence GDGTF (SEQ ID NO:920) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., D replaced by E, T replaced by S, or F replaced by L, W, or Y).

[0150] With regard to FIG.8A, in some cases: i) aac1 (amino acid cluster 1) may be the amino acid sequence RNLRG (SEQ ID NO:921) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., N replaced by T or I; and / or L replaced by A; and / or the second R replaced by L; and / or the G replaced by R); ii) aac2 (amino acid cluster 2) may be the amino acid sequence YNQSE (SEQ ID NO:916) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., N replaced by Q, Q replaced by N, and / or E replaced by D); iii) aac3 (amino acid cluster 3) may be the amino acid sequence TAADT (SEQ ID NO:922) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., the first T replaced by S; and / or A replaced by G; and / or D replaced by E; and / or the second T replaced by S); iv) aac4 (amino acid cluster 4) may be the amino acid sequence AQITQ (SEQ ID NO:923) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., A replaced by G; and / or the first Q replaced by N; and / or I replaced by L or V; and / or the T replaced by S; and / or the second Q replaced by N); v) aac5 (amino acid cluster 5) may be the amino acid sequence VETRP (SEQ ID NO:919) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., V replaced by I or L, E replaced by D, T replaced by S, and / or R replaced by K); and / or vi) aac6 (amino acid cluster 6) may be the amino acid sequence GDRTF (SEQ ID NO:924) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., D replaced by E; and / or T replaced by S; and / or R replaced by K or H; and / or F replaced by L, W, or Y).

[0151] With regard to FIG.9A, in some cases: i) aac1 (amino acid cluster 1) may be the amino acid sequence RNLRG (SEQ ID NO:921) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., N replaced by K; and / or L replaced by A or I; and / or the second R replaced by H; and / or the G replaced by T or S); ii) aac2 (amino acid cluster 2) may be the amino acid sequence YNQSE (SEQ ID NO:916) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., N replaced by Q, Q replaced by N, and / or E replaced by D); iii) aac3 (amino acid cluster 3) may be the amino acid sequence TAADT (SEQ ID NO:922) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., the first T replaced by S; and / or A replaced by G; and / or D replaced by E; and / or the second T replaced by S); iv) aac4 (amino acid cluster 4) may be the amino acid sequence AQITQ (SEQ ID NO:923) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., A replaced by G; and / or the first Q replaced by N; and / or I replaced by L; and / or the second Q replaced by N or K); v) aac5 (amino acid cluster 5) may be the amino acid sequence VETRP (SEQ ID NO:919) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., V replaced by I or L, E replaced by D, T replaced by S, and / or R replaced by K or H); and / or vi) aac6 (amino acid cluster 6) may be theamino acid sequence GDGTF (SEQ ID NO:920) or that sequence with one or two amino acids deleted or substituted with other naturally occurring amino acids (e.g., D replaced by E; and / or T replaced by S; and / or F replaced by L, W, or Y). Non-classical HLA-E, -F, and -G MHC class I heavy chains

[0152] In some cases, a TMMP comprises a non-classical MHC class I heavy chain polypeptide. Among the non-classical HLA heavy chain polypeptides, or portions thereof, that may be that may be incorporated into a TMMP include, but are not limited to, those of HLA-E, -F, and -G alleles. Amino acid sequences for HLA-E, -F, and -G heavy chain polypeptides, (and the HLA-A, B and C alleles) may be found on the world wide web hla.alleles.org / nomenclature / index.html, the European Bioinformatics Institute (www(dot)ebi(dot)ac(dot)uk), which is part of the European Molecular Biology Laboratory(EMBL), and at the National Center for Biotechnology Information (www(dot)ncbi(dot)nlm(dot)nih(dot)gov).

[0153] Non-limiting examples of suitable HLA-E alleles include, but are not limited to, HLA- E*0101 (HLA-E*01:01:01:01), HLA-E*01:03(HLA-E*01:03:01:01), HLA-E*01:04, HLA-E*01:05, HLA-E*01:06, HLA-E*01:07, HLA-E*01:09, and HLA-E*01:10. Of these, isoforms HLA-E*0101 and HLA-E*01.03 are of particular note since these are highly prevalent alleles, and differ by only 1 amino acid (Arg or Gly at position 107). For example, amino acid sequences of suitable HLA-E heavy chain polypeptides are provided in FIG.16A-16D, where FIG.16A provides the amino acid sequence of HLA-E*01:01 (wild-type); FIG.16B provides the amino acid sequence of HLA-E*01:01 with Y84C and A2346C substitutions; FIG.16C provides the amino acid sequence of HLA-E*01:03 (wild-type); and FIG.16D provides the amino acid sequence of HLA-E*01:03 with Y84C and A2346C substitutions. In some cases, therefore, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.16A, 16B, 16C or 16D.

[0154] Non-limiting examples of suitable HLA-F alleles include, but are not limited to, HLA- F*0101 (HLA-F*01:01:01:01), HLA-F*01:02, HLA-F*01:03(HLA-F*01:03:01:01), HLA-F*01:04, HLA-F*01:05, and HLA-F*01:06. Non-limiting examples of suitable HLA-G alleles include, but are not limited to, HLA-G*0101 (HLA-G*01:01:01:01), HLA-G*01:02, HLA-G*01:03(HLA-G*01:03:01:01), HLA-G*01:04 (HLA-G*01:04:01:01), HLA-G*01:06, HLA-G*01:07, HLA-G*01:08, HLA-G*01:09: HLA-G*01:10, HLA-G*01:10, HLA-G*01:11, HLA-G*01:12, HLA-G*01:14, HLA-G*01:15, HLA- G*01:16, HLA-G*01:17, HLA-G*01:18: HLA-G*01:19, HLA-G*01:20, and HLA-G*01:22. Of these, isoforms HLA-G*0101 (HLA-G*01:01:01:01) and HLA-G*01:04 (HLA-G*01:04:01:01) are of particular note since these are highly prevalent alleles. For example, amino acid sequences of suitableHLA-G heavy chain polypeptides are provided in FIG.17A-17D, where FIG.17A provides the amino acid sequence of HLA-G*01:01 (wild-type); FIG.17B provides the amino acid sequence of HLA- G*01:01 with Y84C and A2346C substitutions; FIG. 17C provides the amino acid sequence of HLA- G*01:04 (wild-type); and FIG.17D provides the amino acid sequence of HLA-G*01:04 with Y84C and A2346C substitutions. In some cases, therefore, an MHC class I heavy chain polypeptide of a TMMP can comprise an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.17A, 17B, 17C or 17D.

[0155] Consensus sequences for those HLA E, -F and -G alleles without all, or substantially all, of the leader, transmembrane and cytoplasmic sequences are provided in FIG.10, and aligned with consensus sequences of the above-mentioned HLA-A, -B and -C alleles in FIG. 11.

[0156] FIG.10 provides a consensus sequence for each of HLA-E, -F, and -G with the variable aa positions indicated as “X” residues sequentially numbered and the locations of aas 84, 139 and 236 double underlined.

[0157] FIG.11 provides an alignment of the consensus amino acid sequences for HLA-A, -B, - C, -E, -F, and -G, which are given in FIGs.7-11. Variable residues in each sequence are listed as “X” with the sequential numbering removed. The locations of aas 84, 139 and 236 are indicated with their flanking five-amino acid clusters that may be replaced by 1 to 5 amino acids selected independently from (i) any naturally occurring amino acid or (ii) any naturally occurring amino acid except proline or glycine are also shown.

[0158] Any of the above-mentioned HLA-E, -F, and / or -G alleles may comprise a substitution at one or more of positions 84, 139 and / or 236 as shown in FIG.11 for the consensus sequences. In some cases, the substitutions may be selected from a: position 84 tyrosine to alanine (Y84A) or cysteine (Y84C), or, in the case of HLA-F, an R84A or R84C substitution; a position 139 alanine to cysteine (A139C), or, in the case of HLA-F, a V139C; and an alanine to cysteine substitution at position 236 (A236C). In addition, an HLA-E, -F and / or -G sequence having at least 75% (e.g., at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%) or 100% amino acid sequence identity to all or part (e.g., 50, 75, 100, 150, 200, or 250 contiguous amino acids) of their respective consensus sequences set forth in FIG.11 may also be employed (e.g., the sequences may comprise 1-25, 1-5, 5-10, 10-15, 15-20, 20-25, or 25-30 amino acid insertions, deletions, and / or substitutions in addition to changes at variable residues listed therein). Beta-2 Microglobulin

[0159] A β2-microglobulin (β2M) polypeptide of a TMMP can be a human β2M polypeptide, a non-human primate β2M polypeptide, a murine β2M polypeptide, and the like. In some instances, a β2Mpolypeptide comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.1A (wild-type human β2M). In some cases, a β2M polypeptide present in a TMMP comprises the amino acid sequence: depicted in FIG.1A (wild-type human β2M).

[0160] In some cases, an MHC polypeptide present in a TMMP comprises a single amino acid substitution relative to a reference MHC polypeptide (where a reference MHC polypeptide can be a wild- type MHC polypeptide), where the single amino acid substitution substitutes an amino acid with a cysteine (Cys) residue. Such cysteine residues can form a disulfide bond with a naturally occurring or non-naturally occurring cysteine residue present in the MHC heavy chain of the TMMP. As used herein, a reference to a “non-naturally occurring Cys residue” in an MHC class I polypeptide means that the polypeptide comprises a Cys residue in a location where there is no Cys in the corresponding wild-type polypeptide. This can be accomplished through routine protein engineering in which a cysteine is substituted for the amino acid that occurs in the wild-type sequence.

[0161] In some cases, a β2M polypeptide present in a TMMP comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.1B, where amino acid 12 is a Cys; i.e., where the β2M comprises a non-naturally-occurring Cys at position 12 as a result of an R12C substitution. In some cases, a β2M polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.1B. IMMUNOMODULATORY POLYPEPTIDES

[0162] In some cases, an immunomodulatory polypeptide (“MOD”) present in a TMMP is a wild-type (“wt”) MOD. As discussed above, in other cases, a MOD present in a TMMP is a variant of a wt. MOD that has reduced affinity for a co-MOD compared to the affinity of a corresponding wild-type MOD for the co-MOD. Suitable MODs that exhibit reduced affinity for a co-MOD can have from 1 amino acid (aa) to 20 aa differences from a wild-type MOD.

[0163] As discussed above, a MOD may comprise a variant of a wt immunomodulatory polypeptide that may exhibit reduced binding to its co-MOD, including e.g., reduced binding to one or more chains or domains of the co-MOD. For example, a variant MOD present in a TMMP may bind its co-MOD with an affinity that it at least 10% less, at least 15% less, at least 20% less, at least 25% less, at least 30% less, at least 35% less, at least 40% less, at least 45% less, at least 50% less, at least 55% less, at least 60% less, at least 65% less, at least 70% less, at least 75% less, at least 80% less, at least 85% less, at least 90% less, at least 95% less, or more than 95% less, than the affinity of a corresponding wild- type MOD for the co-MOD.

[0164] Exemplary pairs of MODs and their co-MODs include, but are not limited to those set out in Table 2, below: Table 2

[0165] One or more MODs can be present in a TMMP at any of a variety of positions. For example, in some cases, a MOD (or multiple MODs) can be: 1) C-terminal to the MHC class I heavy chain and N-terminal to the Ig Fc polypeptide; in other words, between the MHC class I heavy chain polypeptide and the Ig Fc polypeptide; 2) C-terminal to the Ig Fc polypeptide; or 3) N-terminal to the peptide epitope.

[0166] Immunomodulatory polypeptides and variants, including reduced affinity variants, such as PD-L1, CD80, CD86, 4-1BBL and IL-2 are described in the published literature, e.g., published PCT application WO2020132138A1 and WO2019 / 051091, the disclosures of which as they pertain to MODs and specific variant MODs of PD-L1, CD80, CD86, 4-1BBL, IL-2 are expressly incorporated herein byreference, including specifically paragraphs

[0260] -

[0455] of WO2020132138A1 and paragraphs

[0157] -

[0352] of WO2019 / 051091.

[0167] Of specific interest are MODs that are variants of the cytokine IL-2. Wild-type IL-2 binds to IL-2 receptor (IL-2R) on the surface of a T cell. Wild-type IL-2 has a strong affinity for IL-2R and will bind to activate most or substantially all CD8+ T cells. For this reason, synthetic forms of wild type IL-2 such as the drug Aldesleukin (trade name Proleukin®) are known to have severe side-effects when administered to humans for the treatment of cancer because the IL-2 indiscriminately activates both target and non-target T cells.

[0168] An IL-2 receptor is in some cases a heterotrimeric polypeptide comprising an alpha chain (IL-2Rα; also referred to as CD25), a beta chain (IL-2Rβ; also referred to as CD122: and a gamma chain (IL-2Rγ; also referred to as CD132). Amino acid sequences of human IL-2, human IL-2Rα, IL2Rβ, and IL-2Rγ are known. See, e.g., published PCT applications WO2020132138A1 and WO2019 / 051091, discussed above. For example, a wild-type IL-2 polypeptide can have the amino acid sequence depicted in FIG. 12A. Amino acid sequences of human IL-2Rα, human IL-2Rβ, and human IL-2Rγ are depicted in FIG. 12B, FIG.12C, and FIG.12D, respectively. In some cases, the human IL-2Rα, human IL-2Rβ, and human IL-2Rγ polypeptides are the mature form (lacking the signal peptide).

[0169] In some cases, an IL-2 variant MOD of this disclosure exhibits decreased binding to IL- 2Rα, thereby minimizing or substantially reducing the activation of Tregs by the IL-2 variant. Alternatively, or additionally, in some cases, an IL-2 variant MOD of this disclosure exhibits decreased binding to IL-2Rβ such that the IL-2 variant MOD exhibits an overall reduced affinity for IL-2R. In some cases, an IL-2 variant MOD of this disclosure exhibits both properties, i.e., it exhibits decreased or substantially no binding to IL-2Rα, and also exhibits decreased binding to IL-2Rβ such that the IL-2 variant polypeptide exhibits an overall reduced affinity for IL-2R. For example, IL-2 variants having substitutions at H16 and F42 have shown decreased binding to IL-2Rα and IL-2Rβ. See, Quayle et al., Clin Cancer Res; 26(8) April 15, 2020, which discloses that the binding affinity of an IL-2 polypeptide with H16A and F42A substitutions for human IL-2Rα and IL-2Rβ was decreased 110- and 3-fold, respectively, compared with wild-type IL2 binding, predominantly due to a faster off-rate for each of these interactions. TMMPs comprising such variants, including variants that exhibit decreased binding to IL-2Rα and IL-2Rβ, have shown the ability to preferentially bind to and activate IL-2 receptors on T cells that contain the target TCR that is specific for the peptide epitope on the TMMP, and are thus less likely to deliver IL-2 to non-target T cells, i.e., T cells that do not contain a TCR that specifically binds the peptide epitope on the TMMP. That is, the binding of the IL-2 variant MOD to its costimulatory polypeptide on the T cell is substantially driven by the binding of the MHC-epitope moiety rather than by the binding of the IL-2. In some cases, an IL-2 variant MOD of this disclosure exhibits decreasedbinding to IL-2Rγ. This decreased binding to IL-2Rγ may be in addition to the decreased binding to IL- 2Rα and / or IL-2Rβ.

[0170] Suitable IL-2 variant MODs thus include a polypeptide that comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99% amino acid sequence identity to the wild-type IL-2 amino acid sequence depicted in FIG.12A; and that have one or more amino acid differences from the wild-type IL-2 amino acid sequence depicted in FIG. 12A that provide for reduced binding to IL-2Rα, IL-2Rβ and / or IL-2Rγ. In some cases, such a variant IL-2 polypeptide of this disclosure exhibits reduced binding affinity to IL-2R, compared to the binding affinity of an IL-2 polypeptide comprising the wild-type IL-2 amino acid sequence depicted in FIG.12A. For example, in some cases, a variant IL-2 polypeptide binds IL-2R with a binding affinity that is at least 10% less, at least 15% less, at least 20% less, at least 25%, at least 30% less, at least 35% less, at least 40% less, at least 45% less, at least 50% less, at least 55% less, at least 60% less, at least 65% less, at least 70% less, at least 75% less, at least 80% less, at least 85% less, at least 90% less, at least 95% less, or more than 95% less, than the binding affinity of an IL-2 polypeptide comprising the wild-type IL-2 amino acid sequence depicted in FIG.12A for an IL-2R (e.g., an IL-2R comprising polypeptides comprising the amino acid sequences depicted in FIG.12B-12D or comprising the mature forms of the amino acid sequences depicted in FIG.12B-12D), when assayed under the same conditions.

[0171] Mutations that can reduce binding of IL-2 to IL-2Rα include substitutions at one or more of amino acids R38, F42, K43, Y45, E62, P65, E68, V69, and L72. For example, an IL-2 variant can comprise substitutions at one, two, three, four, five or more of the foregoing amino acids. Exemplary substitutions include the following from Table 3: Table 3

[0172] As noted above, a substitution of F42 with an amino acid other than Phe, e.g., Ala, has been shown to substantially reduce the binding of an IL-2 variant to IL-2Rα. See Quayle et al., Clin Cancer Res 2020;26:1953–64, which reported that an F42A substitution caused a 110-fold decrease in the binding to IL-2Rα. The binding to IL-2Rα is thus substantially abrogated by this mutation.

[0173] Mutations that can reduce binding of IL-2 to IL-2Rβ include substitutions at one or more of amino acids E15, H16, L19, D20, D84, S87, N88, V91, I92. For example, an IL-2 variant can comprise substitutions at one, two, three, four, five or more of the foregoing amino acids. Exemplary substitutions include the following from Table 4: Table 4

[0174] As noted above, a substitution of H16 with an amino acid other than His, e.g., Ala, has been shown to reduce the binding of an IL-2 variant to IL-2Rβ. See Quayle et al., Clin Cancer Res 2020;26:1953–64, which reported that an H16A substitution caused a 3-fold decrease in the binding to IL-2Rβ.

[0175] Some exemplary combinations of mutations that reduce binding of an IL-2 variant polypeptide to IL-2Rα and IL-2Rβ include the following from Table 5: Table 5

[0176] Exemplary IL-2 variant polypeptide amino acid sequences comprise an amino acid sequence having at least about least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to any one of the following IL-2 variant amino acid sequences (I) – (XVIII):

[0177] (I) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTX1ML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1225), wherein X1 is any amino acid other than Arg, e.g., Ala, Asp or Glu;

[0178] (II) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TX1KFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1226), wherein X1 is any amino acid other than Phe, e.g., Ala or Lys, and in some cases, X1 instead may be Gly, Val, Leu, Ile, Arg, Asn, Cys, Glu, Gln, Ile, Lys, Met, Phe, Pro, Ser, Tyr, Trp or Val;

[0179] (III) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFX1FYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1227), wherein X1 is any amino acid other than Lys, e.g., Glu;

[0180] (IV) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFX1MPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1228), wherein X1 is any amino acid other than Tyr;

[0181] (V) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EX1LKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1229), wherein X1is any amino acid other than Glu, e.g., Gln;

[0182] (VI) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKX1LEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1230), wherein X1is any amino acid other than Pro;

[0183] (VII) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELPKLEX1VL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1231), wherein X1 is any amino acid other than Glu;

[0184] (VIII) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELPKLEEX1L NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1232), wherein X1 is any amino acid other than Val;

[0185] (IX) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELPKLEEVL NX1AQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1233), wherein X1 is any amino acid other than Leu;

[0186] (X) APTSSSTKKT QLQLX1HLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1234), wherein X1 is any amino acid other than Glu, e.g., Ala;

[0187] (XI) APTSSSTKKT QLQLEX1LLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1235), wherein X1 is any amino acid other than His, e.g., Ala, Glu, Thr, or Asp, and in some cases, X1 is Gly, Val, Leu, Ile, Arg, Asn, Cys, Glu, Gln, Ile, Lys, Met, Phe, Pro, Ser, Tyr, Trp or Val;

[0188] (XII) APTSSSTKKT QLQLEHLLX1D LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1236), wherein X1 is any amino acid other than Leu;

[0189] (XIII) APTSSSTKKT QLQLEHLLLX1LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1237), wherein X1is any amino acid other than Asp, e.g., Asn;

[0190] (XIV) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRX1LISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1238), wherein X1is any amino acid other than Asp, e.g., His, Lys or Arg;

[0191] (XV) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLI X1NIN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1239), wherein X1 is any amino acid other than Ser;

[0192] (XVI) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISX1IN VIVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1240), wherein X1 is any amino acid other than Asn, e.g., Ser, Ala, Gly, Arg, Thr or Asp;

[0193] (XVII) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN X1IVLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1241), wherein X1 is any amino acid other than Val, e.g., Glu, Ala or Thr; and

[0194] (XVIII) APTSSSTKKT QLQLEHLLLD LQMILNGINN YKNPKLTRML TFKFYMPKKA TELKHLQCLE EELKPLEEVL NLAQSKNFHL RPRDLISNIN VX1VLELKGSE TTFMCEYADE TATIVEFLNR WITFCQSIIS TLT (SEQ ID NO:1242), wherein X1 is any amino acid other than Ile, e.g., Ala.

[0195] In any of the foregoing IL-2 variant amino acid sequences (I) – (XVIII), the amino acid sequence may comprise one or more additional mutations that reduce the binding of the IL-2 variant to IL-2Rα and / or IL-2Rβ (as compared to the binding affinity of wild-type IL-2 to IL-2Rα and / or IL-2Rβ).

[0196] The determination of binding affinity may be carried out by known methods.

[0197] In some cases, a suitable variant IL-2 polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100% amino acid sequence identity to the amino acid sequence: APTSSSTKKT QLQLEX1LLLD LQMILNGINN YKNPKLTRML TX2KFYMPKKA TELKHLQCLEEELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:1243), i.e., the variant IL-2 polypeptide has the amino acid sequence of wild-type IL-2 but with H16 and F42 substitutions (X1and X2, shown in bold). As noted above, the histidine at position 16 can be substituted with, e.g., Ala, Glu, Thr, or Asp, but any of the following substitutions also may be employed: Gly, Val, Leu, Ile, Arg, Asn, Cys, Glu, Gln, Ile, Lys, Met, Phe, Pro, Ser, Tyr, or Trp. The phenylalanine at position 42 can be substituted with, e.g., Ala or Lys, but any of the following substitutions also may be employed: Met, Pro, Ser, Thr, Trp, Tyr, or Val.

[0198] In some cases, a variant IL-2 polypeptide present in a TMMP comprises the amino acid sequence: APTSSSTKKT QLQLEALLLD LQMILNGINN YKNPKLTRML TAKFYMPKKA TELKHLQCLEEELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:878).

[0199] In some cases, a variant IL-2 polypeptide present in a TMMP comprises the amino acid sequence: APTSSSTKKT QLQLETLLLD LQMILNGINN YKNPKLTRML TAKFYMPKKA TELKHLQCLEEELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:925). In some cases, a TMMP comprises two copies of such a variant IL-2 polypeptide.

[0200] In some cases, a variant IL-2 polypeptide present in a TMMP comprises the amino acid sequence: APTSSSTKKT QLQLEELLLD LQMILNGINN YKNPKLTRML TAKFYMPKKA TELKHLQCLEEELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:1244). In some cases, a TMMP comprises two copies of such a variant IL-2 polypeptide.

[0201] In some cases, a variant IL-2 polypeptide present in a TMMP comprises the amino acid sequence: APTSSSTKKT QLQLEDLLLD LQMILNGINN YKNPKLTRML TAKFYMPKKA TELKHLQCLEEELKPLEEVL NLAQSKNFHL RPRDLISNIN VIVLELKGSE TTFMCEYADE TATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:1245). In some cases, a TMMP comprises two copies of such a variant IL-2 polypeptide.

[0202] As further examples of MODS, in some cases, a MOD present in a TMMP is a 4-1BBL polypeptide. In some cases, a 4-1BBL polypeptide of a TMMP comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following 4-1BBL amino acid sequence: DPAGLLDLRQG MFAQLVAQNV LLIDGPLSWY SDPGLAGVSL TGGLSYKEDT KELVVAKAGV YYVFFQLELR RVVAGEGSGS VSLALHLQPL RSAAGAAALA LTVDLPPASS EARNSAFGFQ GRLLHLSAGQ RLGVHLHTEA RARHAWQLTQ GATVLGLFRV TPEIPA (SEQ ID NO:926).

[0203] In some cases, a MOD present in a TMMP is a CD80 polypeptide. In some cases, a CD80 polypeptide of a TMMP comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to following CD80 amino acid sequence: VIHVTK EVKEVATLSC GHNVSVEELA QTRIYWQKEK KMVLTMMSGD MNIWPEYKNR TIFDITNNLS IVILALRPSD EGTYECVVLK YEKDAFKREH LAEVTLSVKA DFPTPSISDF EIPTSNIRRI ICSTSGGFPE PHLSWLENGE ELNAINTTVS QDPETELYAV SSKLDFNMTT NHSFMCLIKY GHLRVNQTFN WNTTKQEHFP DN (SEQ ID NO: (SEQ ID NO:927).

[0204] In some cases, a MOD present in a TMMP is a CD86 polypeptide. In some cases, a CD86 polypeptide of a TMMP comprises an amino acid sequence having at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following CD86 amino acid sequence: APLKIQAYFNETADLPCQFANSQNQSLSELVVFWQDQENLVLNEVYLGKEKFDSVHSKYMNRT SFDSDSWTLRLHNLQIKDKGLYQCIIHHKKPTGMIRIHQMNSELSVLANFSQPEIVPISNITENVYI NLTCSSIHGYPEPKKMSVLLRTKNSTIEYDGIMQKSQDNVTELYDVSISLSVSFPDVTSNMTIFCI LETDKTRLLSSPFSIELEDPQPPPDHIP (SEQ ID NO: (SEQ ID NO:928). Anti-CD28 polypeptides

[0205] In some cases, an immunomodulatory polypeptide present in a TMMP is an anti-CD28 antibody (e.g., an anti-CD28 scFv; an anti-CD28 nanobody; an anti-CD28 diabody; and the like). The ability of anti-CD28 antibodies to act as a superagonist, agonist, or antagonist of CD28 activity has been described. See e.g., Poirier et al., (2012) Amer. J. of Transplantation, “CD28-Specific Immunomodulating Antibodies: What Can Be Learned From Experimental Models?” 12:1682-1690. Of particular interest are anti-CD28 antibodies that act as an agonist or superagonist.

[0206] Thus, in some cases, a TMMP comprises an anti-CD28 antibody as at least one of the one or more immunomodulatory polypeptides. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) one or more IL-2 polypeptides. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) one or more IL-2 polypeptides, where the one or more IL-2 polypeptide are variant IL-2 polypeptides, as described above (e.g., an IL-2 polypeptide comprising H16A and F42A substitutions; an IL-2 polypeptide comprising H16T and F42A substitutions; an IL-2 polypeptide comprising H16T and F42T substitutions; and IL-2 polypeptide comprising H16A and F42T substitutions; and the like). In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) two copies of an IL-2 polypeptide comprising H16A and F42A substitutions. Examples of anti-CD28 VL and VH polypeptides are provided in below. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) two copies of an IL-2 polypeptide comprising H16T and F42A substitutions. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) one, two, or three copies of a 4-1BBL polypeptide. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) three copies of a 4-1BBL polypeptide comprising a wild- type amino acid sequence. In some cases, a TMMP comprises, as the one or more immunomodulatory polypeptides: a) an anti-CD28 antibody; and b) three copies of a variant 4-1BBL polypeptide. The two different immunomodulatory polypeptides can be at any of a number of different positions in the first or the second polypeptide of the heterodimer of the TMMP. For example, in some cases, the anti-CD28 antibody is at the C-terminus of the first polypeptide; and the second immunomodulatory polypeptide (e.g., an IL-2 polypeptide (e.g., a variant IL-2 polypeptide); a 4-1BBL polypeptide; etc.) is at the N- terminus of the second polypeptide. As another example, in some cases, the anti-CD28 antibody is at the N-terminus of the second polypeptide; and the second immunomodulatory polypeptide (e.g., an IL-2 polypeptide (e.g., a variant IL-2 polypeptide); a 4-1BBL polypeptide; etc.) is at the C-terminus of the second polypeptide.

[0207] In some cases, an anti-CD28 antibody suitable for inclusion as an immunomodulatory polypeptide in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising the following amino acid sequence: QWYQQKPGQPPKLLIFAASNVESGVPARFSGSGSGTNFSLNIHPVDEDDVAMYFCQQSRKVPYT FGGGTKLEIKR (SEQ ID NO:929); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVKLQQSGPGLVTPSQSLSITCTVSGFSLSDYGVHWVRQSPGQGLEWLGVIWAGGGTNYNSAL MSRKSISKDNSKSQVFLKMNSLQADDTAVYYCARDKGYSYYYSMDYWGQGTTVTVSS (SEQ ID NO:930). In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; andKabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987). In some cases, the VH CDRs are: DYGVH (SEQ ID NO:931) (VH CDR1); VIWAGGGTNYNSALMS (SEQ ID NO:932) (VH CDR2); and DKGYSYYYSMDY (SEQ ID NO:933) (VH CDR3).

[0208] In some cases, an anti-CD28 antibody suitable for inclusion as an immunomodulatory polypeptide in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QWYQQKPGQPPKLLIFAASNVESGVPARFSGSGSGTNFSLNIHPVDEDDVAMYFCQQSRKVPYT FGGGTKLEIKR (SEQ ID NO:929); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVKLQQSGPGLVTPSQSLSITCTVSGFSLSDYGVHWVRQSPGQGLEWLGVIWAGGGTNYNSAL MSRKSISKDNSKSQVFLKMNSLQADDTAVYYCARDKGYSYYYSMDYWGQGTTVTVSS (SEQ ID NO:930).

[0209] In some cases, an anti-CD28 antibody suitable for inclusion as an immunomodulatory polypeptide in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QWYQQKPGQPPKLLIFAASNVESGVPARFSGSGSGTNFSLNIHPVDEDDVAMYFCQQSRKVPYT FGGGTKLEIKR (SEQ ID NO:929); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVKLQQSGPGLVTPSQSLSITCTVSGFSLSDYGVHWVRQSPGQGLEWLGVIWAGGGTNYNSAL MSRKSISKDNSKSQVFLKMNSLQADDTAVYYCARDKGYSYYYSMDYWGQGTTVTVSS (SEQ ID NO:930). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0210] In some cases, an anti-CD28 antibody suitable for inclusion as an immunomodulatory polypeptide in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVKLQQSGPGLVTPSQSLSITCTVSGFSLSDYGVHWVRQSPGQGLEWLGVIWAGGGTNYNSAL MSRKSISKDNSKSQVFLKMNSLQADDTAVYYCARDKGYSYYYSMDYWGQGTTVTVSS (SEQID NO:930); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QWYQQKPGQPPKLLIFAASNVESGVPARFSGSGSGTNFSLNIHPVDEDDVAMYFCQQSRKVPYT FGGGTKLEIKR (SEQ ID NO:929). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids. Immunomodulatory polypeptide and TTP positions

[0211] An immunomodulatory polypeptide (i.e., one or more immunomodulatory polypeptides) can be present in a TMMP at any of a variety of positions. For example, an immunomodulatory polypeptide can be: 1) N-terminal to the MHC class I heavy chain (position 1); 2) C-terminal to the MHC class I heavy chain and N-terminal to the Ig Fc polypeptide; in other words, between the MHC class I heavy chain and the Ig Fc polypeptide (position 2); 3) C-terminal to the Ig Fc polypeptide (position 3); 4) N-terminal to the peptide epitope (position 4); or 5) C-terminal to the β2M polypeptide (position 5). Similarly, a TTP can be present in a TMMP at any of a variety of positions. Non-limiting examples of the various MOD positions and the various TTP positions are illustrated in FIGS.28A-28C.

[0212] Of the embodiments illustrated in FIGS.28A-28C, configurations where the MOD(s) is / are in Position 1 or Position 3, and where the TTP is in Position 1 or Position 3, e.g., (MOD Position 1; TTP Position 3) and (MOD Position 3; TTP Position 1) may be more commonly employed. As also noted above, in any of the above embodiments, one or more of the individual components (e.g., peptide epitope; β2M polypeptide; TTP; etc.) of the first polypeptide in each of the above TMMP combinations are linked together by one or more linkers (e.g., peptide linkers, as described elsewhere herein). In any of the above embodiments, one or more of the individual components (e.g., MHC class I heavy chain; immunomodulatory polypeptide; Ig Fc polypeptide; etc.) of the second polypeptide in each of the above TMMP combinations are linked together by one or more linkers (e.g., peptide linkers, as described elsewhere herein). SCAFFOLD POLYPEPTIDES

[0213] A TMMP can comprise an Fc polypeptide or can comprise another suitable scaffold polypeptide.

[0214] Suitable scaffold polypeptides include antibody-based scaffold polypeptides and non- antibody-based scaffolds. Non-antibody-based scaffolds include, e.g., albumin, an XTEN (extended recombinant) polypeptide, transferrin, an Fc receptor polypeptide, an elastin-like polypeptide (see, e.g., Hassouneh et al. (2012) Methods Enzymol.502:215; e.g., a polypeptide comprising a pentapeptide repeatunit of (Val-Pro-Gly-X-Gly; SEQ ID NO:935), where X is any amino acid other than proline), an albumin-binding polypeptide, a silk-like polypeptide (see, e.g., Valluzzi et al. (2002) Philos Trans R Soc Lond B Biol Sci.357:165), a silk-elastin-like polypeptide (SELP; see, e.g., Megeed et al. (2002) Adv Drug Deliv Rev.54:1075), and the like. Suitable XTEN polypeptides include, e.g., those disclosed in WO 2009 / 023270, WO 2010 / 091122, WO 2007 / 103515, US 2010 / 0189682, and US 2009 / 0092582; see also Schellenberger et al. (2009) Nat Biotechnol.27:1186). Suitable albumin polypeptides include, e.g., human serum albumin.

[0215] Suitable scaffold polypeptides will in some cases be a half-life extending polypeptides. Thus, in some cases, a suitable scaffold polypeptide increases the in vivo half-life (e.g., the serum half- life) of the TMMP, compared to a control TMMP lacking the scaffold polypeptide. For example, in some cases, a scaffold polypeptide increases the in vivo half-life (e.g., the serum half-life) of the TMMP, compared to a control TMMP lacking the scaffold polypeptide, by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 50%, at least about 2-fold, at least about 2.5- fold, at least about 5-fold, at least about 10-fold, at least about 25-fold, at least about 50-fold, at least about 100-fold, or more than 100-fold. As an example, in some cases, an Fc polypeptide increases the in vivo half-life (e.g., the serum half-life) of the TMMP, compared to a control TMMP lacking the Fc polypeptide, by at least about 10%, at least about 15%, at least about 20%, at least about 25%, at least about 50%, at least about 2-fold, at least about 2.5-fold, at least about 5-fold, at least about 10-fold, at least about 25-fold, at least about 50-fold, at least about 100-fold, or more than 100-fold. Fc polypeptides

[0216] In some cases, a TMMP comprises an Ig Fc polypeptide. An Ig Fc polypeptide is also referred to herein as an “Fc polypeptide.” The Ig Fc polypeptide of a TMMP can be a human IgG1 Fc, a human IgG2 Fc, a human IgG3 Fc, a human IgG4 Fc, etc., or a variant of a wild-type Ig Fc polypeptide. Variants include naturally occurring variants, non-naturally occurring variants, and combinations thereof. For example, the Ig Fc can be a variant of a Fc polypeptide such as a human IgG1 Fc, which variant has a substantially reduced ability to effect complement-dependent cytotoxicity (CDC) or antibody- dependent cell cytotoxicity (ADCC). See, e.g., the variant human IgG1 Fc polypeptides of FIG.2B and FIG.2D).

[0217] In some cases, the Fc polypeptide present in a TMMP comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the Fc amino acid sequence depicted in any one of FIG.2A-2M.

[0218] In some cases, the Fc polypeptide present in a TMMP is an IgG1 Fc polypeptide, or a variant of an IgG1 Fc polypeptide. For example, in some cases, the Fc polypeptide present in a TMMPcomprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG1 Fc polypeptide depicted in FIG.2A. As another example, in some cases, the Fc polypeptide present in a TMMP comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the Fc polypeptide depicted in FIG.2B, where the Ig Fc polypeptide comprises an Ala at position 14 and an Ala at position 15. In any of the above embodiments, the Ig Fc polypeptide can have an N77 substitution, i.e., the Ig Fc polypeptide can have an amino acid other than Asn at position 77, where in some cases, the Ig Fc polypeptide has an Ala at position 77. In some cases, an Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.2A. In some cases, an Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG. 2B.

[0219] In some cases, the Fc polypeptide present in a TMMP is an IgG1 Fc polypeptide, or a variant of an IgG1 Fc polypeptide, where variants include naturally occurring variants, non-naturally- occurring variants, and combinations thereof. For example, in some cases, the Fc polypeptide present in a TMMP comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG1 Fc polypeptide depicted in FIG.2C; where the Ig Fc polypeptide comprises a Glu at position 136 and a Met at position 138. As another example, in some cases, the Fc polypeptide present in a TMMP comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG1 Fc polypeptide depicted in FIG.2D; where the Ig Fc polypeptide has Ala at positions 14 and 15; and where the Fc polypeptide comprises a Glu at position 136 and a Met at position 138. In any of the above embodiments, the Ig Fc polypeptide can have an N77 substitution, i.e., the Ig Fc polypeptide can have an amino acid other than Asn at position 77, where in some cases, the Ig Fc polypeptide has an Ala at position 77. In some cases, an Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.2C. In some cases, an Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.2D.

[0220] In some cases, the Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.2E (human IgG1 Fc comprising an L234F substitution, an L235E substitution, and a P331S substitution; where L234 corresponds to amino acid 14 of the amino acid sequence depicted in FIG.2A; L235 corresponds to amino acid 15 of the amino acid sequence depicted in FIG. 2E; and P331 corresponds to amino acid 111 of the amino acid sequence depicted in FIG.2E). In some cases, the Fc polypeptide present in a TMMP comprises the amino acid sequence depicted in FIG.2F, comprising an N279A substitution (N77A of the amino acid sequence depicted in FIG.2F). Substitutions at N297 lead to the removal of carbohydrate modifications and result antibody sequenceswith reduced complement component 1q (“C1q”) binding compared to the wild-type protein, and accordingly a reduction in complement-dependent cytotoxicity (CDC). In some cases, the Fc polypeptide present in a TMMP comprises a substitution at K322. K322 (e.g., K322A) substitutions shows a substantial reduction in reduction in FcγR binding affinity and a reduction in antibody-dependent cell- mediated cytotoxicity (ADCC), with the C1q binding and CDC functions substantially or completely eliminated.

[0221] In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG2 Fc polypeptide depicted in FIG.2G; e.g., the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to amino acids 99-325 of the human IgG2 Fc polypeptide depicted in FIG.2G (e.g., where the Ig Fc polypeptide has a length of about 227 amino acids). In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG3 Fc polypeptide depicted in FIG.2H; e.g., the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to amino acids 19-246 of the human IgG3 Fc polypeptide depicted in FIG.2H (e.g., where the Ig Fc polypeptide has a length of about 228 amino acids). In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgM Fc polypeptide depicted in FIG.2J; e.g., the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to amino acids 1-276 to the human IgM Fc polypeptide depicted in FIG.2J. In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgA Fc polypeptide depicted in FIG.2K; e.g., the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to amino acids 1-234 to the human IgA Fc polypeptide depicted in FIG. 2K.

[0222] In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the human IgG4 Fc polypeptide depicted in FIG.2M. In some cases, the Fc polypeptide comprises an amino acid sequence having at least 70%, at least 75%, at least80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to amino acids 100 to 327 of the human IgG4 Fc polypeptide depicted in FIG.2M (e.g., where the Ig Fc polypeptide has a length of about 228 amino acids).

[0223] In some cases, the IgG4 Fc polypeptide comprises the following amino acid sequence: PPCPSCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNA KTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTL PPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKS RWQEGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:936). LINKERS

[0224] As discussed above, a TMMP can include one or more peptide linkers, i.e., a linker comprising a contiguous stretch of two or more amino acids, where the one or more linkers are between one or more components of a TMMP. For example, a TMMP can include one or more peptide linkers, i.e., a linker comprising a contiguous stretch of two or more amino acids, where the one or more linkers are between one or more of: i) an MHC class I heavy chain polypeptide and an Ig Fc polypeptide; ii) a MOD and an MHC class I polypeptide; iii) a first MOD and a second MOD; iv) a peptide and an MHC class I polypeptide; v) a peptide and a β2M polypeptide; vi) a TTP and an MHC class I heavy chain polypeptide; vii) a TTP and an Ig Fc polypeptide; and viii) a TTP and a MOD.

[0225] As used herein, the phrase “an optional peptide linker between any two of the components of a TMMP” refers to a peptide linker between any two adjacent polypeptides within the TMMP. For example, as used herein, the phrase “an optional peptide linker between any two of the components of a TMMP” refers to a peptide linker between one or more of: ) an MHC class I heavy chain polypeptide and an Ig Fc polypeptide; ii) a MOD and an MHC class I polypeptide; iii) a first MOD and a second MOD; iv) a peptide and an MHC class I polypeptide; v) a peptide and a β2M polypeptide; vi) a TTP and an MHC class I heavy chain polypeptide; vii) a TTP and an Ig Fc polypeptide; and viii) a TTP and a MOD. As discussed below, linkers may be: a) a flexible peptide linker, including a short flexible peptide linker; or b) a rigid peptide linker.

[0226] Suitable linkers (also referred to as “spacers”) can be readily selected and can be of any of a number of suitable lengths, such as from 1 amino acid to 25 amino acids, from 3 amino acids to 20 amino acids, from 2 amino acids to 15 amino acids, from 3 amino acids to 12 amino acids, including 4 amino acids to 10 amino acids, 5 amino acids to 9 amino acids, 6 amino acids to 8 amino acids, or 7 amino acids to 8 amino acids. A suitable linker can be 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 amino acids in length. In some cases, a linker has a length of from 25 amino acids to 50 amino acids, e.g., from 25 to 30, from 30 to 35, from 35 to 40, from 40 to 45, or from 45 to 50 amino acids in length.Flexible peptide linkers

[0227] Exemplary flexible peptide linkers include glycine polymers (G)n (SEQ ID NO:946), glycine-serine polymers (including, for example, (GS)n (SEQ ID NO:937), (GSGGS)n (SEQ ID NO:938), (GGGGS)n (SEQ ID NO:934), and (GGGS)n (SEQ ID NO:939), where n is an integer of at least one and can be an integer from 1 to 10), glycine-alanine polymers, alanine-serine polymers, and other flexible peptide linkers known in the art. Glycine and glycine-serine polymers can be used; both Gly and Ser are relatively unstructured, and therefore can serve as a neutral tether between components. Glycine polymers can be used; glycine accesses significantly more phi-psi space than even alanine, and is much less restricted than residues with longer side chains (see Scheraga, Rev. Computational Chem. 11173-142 (1992)). Exemplary linkers can comprise amino acid sequences including, but not limited to, GGSG (SEQ ID NO:940), GGSGG (SEQ ID NO:941), GSGSG (SEQ ID NO:942), GSGGG (SEQ ID NO:943), GGGSG (SEQ ID NO:944), GSSSG (SEQ ID NO:945), and the like.

[0228] Exemplary flexible peptide linkers include, e.g., (GGGGS)n (SEQ ID NO:934); also referred to as a “G4S” linker), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some cases, a linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10. In some cases, a linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:947), where n is 2. In some cases, a linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:875), where n is 3. In some cases, a linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:876), where n is 4. In some cases, a linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:948), where n is 7. In some cases, a linker comprises the amino acid sequence AAAGG (SEQ ID NO:874). Also suitable is a linker having the amino acid sequence AAAGG (SEQ ID NO:874). In TMMPs of this disclosure, the β2M polypeptide can be connected to the MHC heavy chain polypeptide by a (GGGGS)n (SEQ ID NO:934) linker, where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., where n=3, n=4, or n=7.

[0229] As used in this disclosure, a “short flexible peptide linker” means a flexible peptide linker that comprises fewer than 15 amino acids, i.e., from 2-14 amino acids. For example, a short flexible peptide linker can comprise from 2-4 amino acids, from 2-5 amino acids, from 3-6 amino acids, from 4-8 amino acids, from 5-10 amino acids, or from 10-14 amino acids. Within this range includes flexible peptide linkers comprising 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 amino acids. Rigid peptide linkers

[0230] In some cases, a peptide linker is a rigid peptide linker. As used herein, the term “rigid peptide linker” refers to a linker comprising a contiguous stretch of two or more amino acids that effectively separates protein domains by maintaining a substantially fixed distance / spatial separation between the domains, thereby reducing or substantially eliminating unfavorable interactions between such domains. For example, rigid linkers may be interposed when either the MOD(s) or the TTP are in Position 1 and / or in Position 3 discussed above. Rigid peptide linkers are known in the art and generallyadopt a relatively well-defined conformation when in solution. Rigid peptide linkers include those which have a particular secondary and / or tertiary structure in solution; and are typically of a length sufficient to confer secondary or tertiary structure to the linker. Rigid peptide linkers include peptide linkers rich in proline, and peptide linkers having an inflexible helical structure, such as an α-helical structure. Rigid peptide linkers are described in, for example, Chen et al. (2013) Adv. Drug Deliv. Rev.65:1357; and Klein et al. (2014) Protein Engineering, Design & Selection 27:325.

[0231] Examples of rigid peptide linkers include, e.g., (EAAAK)n (SEQ ID NO:949), A(EAAAK)nA (SEQ ID NO:950), A(EAAAK)nALEA(EAAAK)nA (SEQ ID NO:951), (Lys-Pro)n (SEQ ID NO:952), (Glu-Pro)n (SEQ ID NO:953), (Thr-Pro-Arg)n (SEQ ID NO:954), and (Ala-Pro)n (SEQ ID NO:955) where n is an integer from 1 to 20 (e.g., n is 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20). Non-limiting examples of suitable rigid peptide linkers comprising EAAAK (SEQ ID NO:956) include EAAAK (SEQ ID NO:956), (EAAAK)2 (SEQ ID NO:958), (EAAAK)3 (SEQ ID NO:959), A(EAAAK)4ALEA(EAAAK)4A (SEQ ID NO:960), and AEAAAKEAAAKA (SEQ ID NO:961). Non-limiting examples of suitable rigid peptide linkers comprising (AP)n include APAP (SEQ ID NO:962; also referred to herein as “(AP)2”); APAPAPAP (SEQ ID NO:963; also referred to herein as “(AP)4”); APAPAPAPAPAP (SEQ ID NO:964; also referred to herein as “(AP)6”); APAPAPAPAPAPAPAP (SEQ ID NO:965; also referred to herein as “(AP)8”); and APAPAPAPAPAPAPAPAPAP (SEQ ID NO:955; also referred to herein as “(AP)10”). Non-limiting examples of suitable rigid peptide linkers comprising (KP)n include KPKP (SEQ ID NO:967; also referred to herein as “(KP)2”); KPKPKPKP (SEQ ID NO:968; also referred to herein as “(KP)4”); KPKPKPKPKPKP (SEQ ID NO:969; also referred to herein as “(KP)6”); KPKPKPKPKPKPKPKP (SEQ ID NO:970; also referred to herein as “(KP)8”); and KPKPKPKPKPKPKPKPKPKP (SEQ ID NO:952; also referred to herein as “(KP)10”). Non-limiting examples of suitable rigid peptide linkers comprising (EP)n include EPEP (SEQ ID NO:972; also referred to herein as “(EP)2”); EPEPEPEP (SEQ ID NO:973; also referred to herein as “(EP)4”); EPEPEPEPEPEP (SEQ ID NO:974; also referred to herein as “(EP)6”); EPEPEPEPEPEPEPEP (SEQ ID NO:975; also referred to herein as “(EP)8”); and EPEPEPEPEPEPEPEPEPEP (SEQ ID NO:953; also referred to herein as “(EP)10”). Cysteine-containing linkers

[0232] As noted above, in some cases, a linker peptide in a first polypeptide chain of a TMMP can include a cysteine residue that can form an interchain disulfide bond with a cysteine residue present in a second polypeptide chain. For example, as discussed above, in some cases a TMMP, or a dimerized TMMP such as a homodimer, comprises a linker between the peptide epitope and the β2M polypeptide that includes a cysteine residue that forms an interchain disulfide bond with a cysteine residue in the MHC class I heavy chain polypeptide present in the other chain of the TMMP. For example, in some cases, where a TMMP, or a dimerized TMMP such as a homodimer, comprises a cysteine-containinglinker between the peptide epitope and the β2M polypeptide, the cysteine residue in the linker forms an interchain disulfide bond with a cysteine residue at amino acid 236 (e.g., formed by an A236C substitution) in the MHC class I heavy chain polypeptide present in the TMMP.

[0233] In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence GCGGS (SEQ ID NO:977). In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence GCGGS(GGGGS)n (SEQ ID NO:978), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; e.g., 1, 2, or 3. In some cases, the peptide linker between the peptide and the β2M polypeptide comprises the amino acid sequence GCGGS(GGGGS)n (SEQ ID NO:882), where n is 2.

[0234] In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence CGGGS (SEQ ID NO:979). In some cases, the peptide linker comprises the amino acid sequence CGGGS(GGGGS)n (SEQ ID NO:980), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10; e.g., 1, 2, or 3.

[0235] In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence GGCGS (SEQ ID NO:981). In some cases, the peptide linker comprises the amino acid sequence GGCGS(GGGGS)n (SEQ ID NO:982), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10, e.g., 1, 2, or 3.

[0236] In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence GGGCS (SEQ ID NO:983). In some cases, the peptide linker comprises the amino acid sequence GGGCS(GGGGS)n (SEQ ID NO:984), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, or 3.

[0237] In some cases, the peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide comprises the amino acid sequence GGGGC (SEQ ID NO:985). In some cases, the peptide linker comprises the amino acid sequence GGGGC(GGGGS)n (SEQ ID NO:986), where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, e.g., 1, 2, or 3. Linkers between the TTP and MHC Class I heavy chain polypeptide As discussed below, it has been found that the cytotoxic effect of TMMPs is greater when a TTP is positioned N-terminal to the MHC heavy chain polypeptide and the linker between them is no more than about 12 amino acids in length, e.g., no more than 10 amino acids in length (e.g., a (GGGGS)2 (SEQ ID NO:947) linker), no more than 8 amino acids in length, no more than 7 amino acids in length, no more than 6 amino acids in length, or no more than 5 amino acids in length, (e.g., a GGGGS (SEQ ID NO:873) linker). In some cases, the linker between them is no more than about 12 amino acids in length. In some cases, the linker between them is no more than 10 amino acids in length (e.g., a (GGGGS)2 (SEQ ID NO:947) linker). In some cases, the linker between them is no more than 8 amino acids inlength. In some cases, the linker between them is no more than 7 amino acids in length. In some cases, the linker between them is no more than 6 amino acids in length. In some cases, the linker between them is no more than 5 amino acids in length, (e.g., a GGGGS (SEQ ID NO:873) linker). DIMERIZED TMMPS

[0238] As noted above, in some cases, a TMMP can form dimers. That is, the present disclosure provides a polypeptide comprising a dimer of two TMMPs. The present disclosure thus provides a protein that is a dimerized TMMP comprising two TMMPs that are covalently linked to each other. The covalent linkage of the dimer can be one or more disulfide bonds between an Ig Fc polypeptide in the first TMMP and an Ig Fc polypeptide in the second TMMP. As but one example, the Ig Fc can be a variant of a human IgG1 Fc polypeptide, which variant has a substantially reduced ability to effect complement-dependent cytotoxicity (CDC) or antibody-dependent cell cytotoxicity (ADCC) (e.g., the human IgG1 Fc polypeptide of FIG.2B or FIG.2D). When the TMMP comprises an Ig Fc polypeptide, the TMMP typically will self-assemble into a dimer by spontaneously forming disulfide bonds with the IgG1 Fc polypeptide of another TMMP. Thus, e.g., the Ig Fc polypeptides in the first TMMP and the second TMMP can be linked to one another by one or more disulfide bonds. In many cases, the two TMMPs will be identical to one another in amino acid sequence and comprise Ig Fc polypeptides that spontaneously form one or more disulfide bonds, thereby forming a dimerized TMMP that is a homodimer.

[0239] Accordingly, the present disclosure provides a protein comprising: a) a first TMMP; and b) a second TMMP, which optionally may be identical to the first TMMP, where the first and second TMMPs are covalently linked to one another. The covalent linkage can be a disulfide bond between an Ig Fc polypeptide in the first TMMP and an Ig Fc polypeptide in the second TMMP.

[0240] If desired, the Ig Fc polypeptides of each TMMP can comprise interspecific dimerization sequences, e.g., “Knob-in-Hole” sequences that permit two different TMMPs to selectively dimerize. Interspecific binding sequences favor formation of heterodimers with their cognate polypeptide sequence (i.e., the interspecific sequence and its counterpart interspecific sequence), particularly those based on Ig Fc sequence variants. Such interspecific polypeptide sequences include Knob-in-Hole, and Knob-in-Hole sequences that facilitate the formation of one or more disulfide bonds. For example, one interspecific binding pair comprises a T366Y and Y407T mutant pair in the CH3 domain interface of IgG1, or the corresponding residues of other immunoglobulins. See Ridgway et al., Protein Engineering 9:7, 617-621 (1996). A second interspecific binding pair involves the formation of a knob by a T366W substitution, and a hole by the triple substitutions T366S, L368A and Y407V on the complementary Ig Fc sequence. See Xu et al. mAbs 7:1, 231-242 (2015). Another interspecific binding pair has a first Fc polypeptide with Y349C, T366S, L368A, and Y407V substitutions and a second Ig Fc polypeptide with S354C, andT366W substitutions (disulfide bonds can form between the Y349C and the S354C). See, e.g., Brinkmann and Konthermann, mAbs 9:2, 182–212 (2015). Ig Fc polypeptide sequences, either with or without knob-in-hole modifications, can be stabilized by the formation of disulfide bonds between the Ig Fc polypeptides (e.g., the hinge region disulfide bonds). Thus, in some cases, a dimerized TMMP can be a heterodimer, comprising two TMMP chains that are not identical in amino acid sequence.

[0241] Interspecific dimerization sequences also may be employed to enable TMMPs to be linked to non-TMMP molecules that can provide additional functionality to the TMMP. For example, a TMMP could be linked to a molecule that comprise polypeptides (e.g., antibodies or binding fragments thereof such as scFvs) that bind to cancer-associated antigens, thereby enabling the TMMP to localize to tissues comprising the cancer-associated antigen. Tumor-targeting polypeptides (TTPs)

[0242] As discussed above, a TMMP includes, in the first and / or the second polypeptide, a tumor-targeting polypeptide (TTP), i.e., a polypeptide specific for a cancer-associated epitope. A “cancer-associated” epitope is an epitope that is present in a cancer-associated antigen. In some cases, a TTP is an antibody. In some cases, a TTP is a single-chain T-cell receptor (scTCR). Targets

[0243] In some cases, a TTP present in a TMMP targets a cancer-associated antigen. In some cases, the target of a TTP is a peptide / HLA (pHLA) complex on the surface of a cancer cell, where the peptide can be a cancer-associated peptide (e.g., a peptide fragment of a cancer-associated antigen). Cancer-associated antigens

[0244] Cancer-associated antigens that can be targeted with a tumor-targeting polypeptide present in a TMMP include, e.g., NY-ESO (New York Esophageal Squamous Cell Carcinoma 1), MART-1 (melanoma antigen recognized by T cells 1, also known as Melan-A), HPV (human papilloma virus) E6, BCMA (B-cell maturation antigen), CD123, CD133, CD171, CD19, CD20, CD22, CD30, CD33, CD38, CD138, CEA (carcinoembryonic antigen), EGFR (epidermal growth factor receptor), EGFRvIII (epidermal growth factor receptor variant III), EpCAM (epithelial cell adhesion molecule), EphA2 (ephrin type-A receptor 2), disialoganglioside GD2, GPC3 (glypican-3), HER2, IL13Ralpha2 (Interleukin 13 receptor subunit alpha-2), LeY (a difucosylated type 2 blood group-related antigen), MAGE-A3 (melanoma-associated antigen 3), melanoma glycoprotein, mesothelin, MUC1 (mucin 1), MUC16 (mucin-16), myelin, NKG2D (Natural Killer Group 2D) ligands, PSMA (prostate specific membrane antigen), and ROR1 (type I receptor tyrosine kinase-like orphan receptor).

[0245] Cancer-associated antigens that can be targeted with a TTP present in a TMMP include, but are not limited to, 17-1A-antigen, alpha-fetoprotein (AFP), alpha-actinin-4, A3, antigen specific for A33 antibody, ART-4, B7, Ba 733, BAGE, bcl-2, bcl-6, BCMA, BrE3-antigen, CA125, CAMEL, CAP-1, carbonic anhydrase IX (CAIX), CASP-8 / m, CCL19, CCL21, CD1, CD1a, CD2, CD3, CD4, CD5, CD8, CD11A, CD14, CD15, CD16, CD18, CD19, CD20, CD21, CD22, CD23, CD25, CD29, CD30, CD32b, CD33, CD37, CD38, CD40, CD40L, CD44, CD45, CD46, CD52, CD54, CD55, CD59, CD64, CD66a-e, CD67, CD70, CD70L, CD74, CD79a, CD79b, CD80, CD83, CD95, CD123, CD126, CD132, CD133, CD138, CD147, CD154, CD171, CDC27, CDK-4 / m, CDKN2A, CEA, CEACAM5, CEACAM6, claudin (e.g., claudin-1, claudin-10, claudin-18 (e.g., claudin-18, isoform 2)), complement factors (such as C3, C3a, C3b, C5a and C5), colon-specific antigen-p (CSAp), c-Met, CTLA-4, CXCR4, CXCR7, CXCL12, DAM, Dickkopf-related protein (DKK), ED-B fibronectin, epidermal growth factor receptor (EGFR), EGFRvIII, EGP-1 (TROP-2), EGP-2, ELF2-M, Ep-CAM, EphA2, EphA3, fibroblast activation protein (FAP), fibroblast growth factor (FGF), Flt-1, Flt-3, folate binding protein, folate receptor, G250 antigen, gangliosides (such as GC2, GD3 and GM2), GAGE, GD2, gp100, GPC3, GRO- 13, HLA-DR, HM1.24, human chorionic gonadotropin (HCG) and its subunits, HER2, HER3, HMGB-1, hypoxia inducible factor (HIF-1), HIF-1a, HSP70-2M, HST-2, Ia, IFN-gamma, IFN-alpha, IFN-beta, IFN-X, IL-4R, IL-6R, IL-13R, IL13Ralpha2, IL-15R, IL-17R, IL-18R, IL-2, IL-6, IL-8, IL-12, IL-15, IL-17, IL-18, IL-23, IL-25, ILGF, ILGF-1R, insulin-like growth factor-1 (IGF-1), IGF-1R, integrin αVβ3, integrin α5β1, KC4-antigen, killer-cell immunoglobulin-like receptor (KIR), Kras, KS-1-antigen, KS1-4, LDR / FUT, Legamma, macrophage migration inhibitory factor (MIF), MAGE, MAGE-3, MART-1, MART-2, mCRP, MCP-1, melanoma glycoprotein, mesothelin, MIP-1A, MIP-1B, MIF, mucins (such as MUC1, MUC2, MUC3, MUC4, MUC5ac, MUC13, MUC16, MUM-1 / 2 and MUM-3), NCA66, NCA95, NCA90, Nectin-4, NY-ESO-1, PAM4 antigen, pancreatic cancer mucin, PD-1, PD-L1, PD-1 receptor, placental growth factor, p53, PLAGL2, prostatic acid phosphatase, PSA, PRAME, PSMA, P1GF, RSS, RANTES, SAGE, 5100, survivin, survivin-2B, T101, TAC, TAG-72, tenascin, Thomson-Friedenreich antigens, Tn antigen, TNF-alpha, tumor necrosis antigens, TRAG-3, TRAIL receptors, vascular endothelial growth factor (VEGF), VEGF receptor (VEGFR) and WT-1.

[0246] In some cases, the cancer-associated antigen is an antigen associated with a hematological cancer. Examples of such antigens include, but are not limited to, BCMA, C5, CD19, CD20, CD22, CD25, CD30, CD33, CD38, CD40, CD45, CD52, CD56, CD66, CD74, CD79a, CD79b, CD80, CD138, CTLA-4, CXCR4, DKK, EphA3, GM2, HLA-DR beta, integrin αVβ3, IGF-R1, IL6, KIR, PD-1, PD-L1, TRAILR1, TRAILR2, transferrin receptor, and VEGF. In some cases, the cancer-associated antigen is an antigen expressed by malignant B cells, such as CD19, CD20, CD22, CD25, CD38, CD40, CD45, CD74, CD80, CTLA-4, IGF-R1, IL6, PD-1, TRAILR2, or VEGF.

[0247] In some cases, the cancer-associated antigen is an antigen associated with a solid tumor. Examples of such antigens include, but are not limited to, CAIX, cadherins, CEA, c-MET, CTLA-4, EGFR family members, EpCAM, EphA3, FAP, folate-binding protein, FR-alpha, gangliosides (such as GC2, GD3 and GM2), HER2, HER3, IGF-1R, integrin αVβ3, integrin α5β1, Legamma, Liv1, mesothelin,mucins, NaPi2b, PD-1, PD-L1, PD-1 receptor, pgA33, PSMA, RANKL, ROR1, TAG-72, tenascin, TRAILR1, TRAILR2, VEGF, VEGFR, and others listed above. Peptide / HLA complexes

[0248] In some cases, the target of a TTP is a peptide / HLA (pHLA) complex on the surface of a cancer cell, where the peptide can be a cancer-associated peptide (e.g., a peptide fragment of a cancer- associated antigen). Cancer-associated peptides are known in the art. In some cases, a cancer-associated peptide is bound to an HLA complex comprising an HLA-A*0201 heavy chain and a β2M polypeptide.

[0249] In some cases, the epitope present in the pHLA on the surface of a cancer cell is bound to an HLA complex comprising an HLA heavy chain such as HLA-A*0101, A*0201, A*0301, A*1101, A*2301, A*2402, A*2407, A*3303, and / or A*3401. In some cases, the epitope present in the pHLA on the surface of a cancer cell is bound to an HLA complex comprising an HLA heavy chain such as HLA- B*0702, B*0801, B*1502, B*3802, B*4001, B*4601, and / or B*5301. In some cases, the epitope present in the pHLA on the surface of a cancer cell is bound to an HLA complex comprising an HLA heavy chain such as C*0102, C*0303, C*0304, C*0401, C*0602, C*0701, C*702, C*0801, and / or C*1502.

[0250] In some cases, the epitope is a cancer-associated epitope of any one of the following cancer-associated antigens: a MUC1 polypeptide, an LMP2 polypeptide, an epidermal growth factor receptor (EGFR) vIII polypeptide, a HER-2 / neu polypeptide, a melanoma antigen family A, 3 (MAGE A3) polypeptide, a p53 polypeptide, a mutant p53 polypeptide, an NY-ESO-1 polypeptide, a folate hydrolase (prostate-specific membrane antigen; PSMA) polypeptide, a carcinoembryonic antigen (CEA) polypeptide, a claudin polypeptide (e.g., claudin-1, claudin-10, claudin-18 (e.g., claudin-18, isoform 2)), a Nectin-4 polypeptide, a melanoma antigen recognized by T-cells (melanA / MART1) polypeptide, a Ras polypeptide, a gp100 polypeptide, a proteinase3 (PR1) polypeptide, a bcr-abl polypeptide, a tyrosinase polypeptide, a survivin polypeptide, a prostate specific antigen (PSA) polypeptide, an hTERT polypeptide, a sarcoma translocation breakpoints polypeptide, a synovial sarcoma X (SSX) breakpoint polypeptide, an EphA2 polypeptide, an acid phosphatase, prostate (PAP) polypeptide, a melanoma inhibitor of apoptosis (ML-IAP) polypeptide, an epithelial cell adhesion molecule (EpCAM) polypeptide, an ERG (TMPRSS2 ETS fusion) polypeptide, a NA17 polypeptide, a paired-box-3 (PAX3) polypeptide, an anaplastic lymphoma kinase (ALK) polypeptide, an androgen receptor polypeptide, a cyclin B1 polypeptide, an N-myc proto-oncogene (MYCN) polypeptide, a Ras homolog gene family member C (RhoC) polypeptide, a tyrosinase-related protein-2 (TRP-2) polypeptide, a mesothelin polypeptide, a prostate stem cell antigen (PSCA) polypeptide, a melanoma associated antigen-1 (MAGE A1) polypeptide, a cytochrome P4501B1 (CYP1B1) polypeptide, a placenta-specific protein 1 (PLAC1) polypeptide, a BORIS polypeptide (also known as CCCTC-binding factor or CTCF), an ETV6-AML polypeptide, a breast cancer antigen NY-BR-1 polypeptide (also referred to as ankyrin repeat domain- containing protein 30A), a regulator of G-protein signaling (RGS5) polypeptide, a squamous cellcarcinoma antigen recognized by T-cells (SART3) polypeptide, a carbonic anhydrase IX polypeptide, a paired box-5 (PAX5) polypeptide, an OY-TES1 (testis antigen; also known as acrosin binding protein) polypeptide, a sperm protein 17 polypeptide, a lymphocyte cell-specific protein-tyrosine kinase (LCK) polypeptide, a high molecular weight melanoma associated antigen (HMW-MAA), an A-kinase anchoring protein-4 (AKAP-4), a synovial sarcoma X breakpoint 2 (SSX2) polypeptide, an X antigen family member 1 (XAGE1) polypeptide, a B7 homolog 3 (B7H3; also known as CD276) polypeptide, a legumain polypeptide (LGMN1; also known as asparaginyl endopeptidase), a tyrosine kinase with Ig and EGF homology domains-2 (Tie-2; also known as angiopoietin-1 receptor) polypeptide, a P antigen family member 4 (PAGE4) polypeptide, a vascular endothelial growth factor receptor 2 (VEGF2) polypeptide, a MAD-CT-1 polypeptide, a fibroblast activation protein (FAP) polypeptide, a platelet derived growth factor receptor beta (PDGFβ) polypeptide, a MAD-CT-2 polypeptide, a Fos-related antigen-1 (FOSL) polypeptide; a human papilloma virus (HPV) antigen; an alpha-feto protein (AFP) antigen; and a Wilms tumor-1 (WT1) antigen.

[0251] For example, in some cases, a TTP present in a TMMP binds to: a) a WT-1 peptide bound to an HLA complex comprising an HLA heavy chain (e.g., an HLA-A*0201 heavy chain or an HLA- A*2402 heavy chain) and a β2M polypeptide; b) an HPV peptide bound to an HLA complex comprising a class I HLA heavy chain and a β2M polypeptide; c) a mesothelin peptide bound to an HLA complex comprising a class I HLA heavy chain and a β2M polypeptide; d) a Her2 peptide bound to an HLA complex comprising a class I HLA heavy chain and a β2M polypeptide; or e) a BCMA peptide bound to an HLA complex comprising a class I HLA heavy chain and a β2M polypeptide.

[0252] In some cases, a cancer-associated peptide is a peptide of a mesothelin polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following mesothelin amino acid sequence:

[0253] LAGE TGQEAAPLDG VLANPPNISS LSPRQLLGFP CAEVSGLSTE RVRELAVALA QKNVKLSTEQ LRCLAHRLSE PPEDLDALPL DLLLFLNPDA FSGPQACTRF FSRITKANVD LLPRGAPERQ RLLPAALACW GVRGSLLSEA DVRALGGLAC DLPGRFVAES AEVLLPRLVS CPGPLDQDQQ EAARAALQGG GPPYGPPSTW SVSTMDALRG LLPVLGQPII RSIPQGIVAA WRQRSSRDPS WRQPERTILR PRFRREVEKT ACPSGKKARE IDESLIFYKK WELEACVDAA LLATQMDRVN AIPFTYEQLD VLKHKLDELY PQGYPESVIQ HLGYLFLKMS PEDIRKWNVT SLETLKALLE VNKGHEMSPQ VATLIDRFVK GRGQLDKDTL DTLTAFYPGY LCSLSPEELS SVPPSSIWAV RPQDLDTCDP RQLDVLYPKA RLAFQNMNGS EYFVKIQSFL GGAPTEDLKA LSQQNVSMDL ATFMKLRTDA VLPLTVAEVQ KLLGPHVEGL KAEERHRPVR DWILRQRQDD LDTLGLGLQG GIPNGYLVLD LSMQEALSGT PCLLGPGPVL TVLALLLAST LA (SEQ ID NO:987). For example, a mesothelin peptide present in a pHLA complex can be: i) KLLGPHVEGL (SEQ ID NO:988); ii) AFYPGYLCSL (SEQ ID NO:989), which can bind HLA-A*2402 / β2M; iii)VLPLTVAEV (SEQ ID NO:990); iv) ELAVALAQK (SEQ ID NO:991); v) ALQGGGPPY (SEQ ID NO:992); vi) FYPGYLCSL (SEQ ID NO:993); vii) LYPKARLAF (SEQ ID NO:994); viii) LLFLLFSLGWVGPSR (SEQ ID NO:995); ix) VNKGHEMSPQAPRRP (SEQ ID NO:996); x) FMKLRTDAVLPLTVA (SEQ ID NO:997); or xi) DAALLATQMD (SEQ ID NO:998).

[0254] In some cases, a cancer-associated peptide is a peptide of a Her2 polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following Her2 (receptor tyrosine-protein kinase erbB2) amino acid sequence:

[0255] MELAALCRWG LLLALLPPGA ASTQVCTGTD MKLRLPASPE THLDMLRHLY QGCQVVQGNL ELTYLPTNAS LSFLQDIQEV QGYVLIAHNQ VRQVPLQRLR IVRGTQLFED NYALAVLDNG DPLNNTTPVT GASPGGLREL QLRSLTEILK GGVLIQRNPQ LCYQDTILWK DIFHKNNQLA LTLIDTNRSR ACHPCSPMCK GSRCWGESSE DCQSLTRTVC AGGCARCKGP LPTDCCHEQC AAGCTGPKHS DCLACLHFNH SGICELHCPA LVTYNTDTFE SMPNPEGRYT FGASCVTACP YNYLSTDVGS CTLVCPLHNQ EVTAEDGTQR CEKCSKPCAR VCYGLGMEHL REVRAVTSAN IQEFAGCKKI FGSLAFLPES FDGDPASNTA PLQPEQLQVF ETLEEITGYL YISAWPDSLP DLSVFQNLQV IRGRILHNGA YSLTLQGLGI SWLGLRSLRE LGSGLALIHH NTHLCFVHTV PWDQLFRNPH QALLHTANRP EDECVGEGLA CHQLCARGHC WGPGPTQCVN CSQFLRGQEC VEECRVLQGL PREYVNARHC LPCHPECQPQ NGSVTCFGPE ADQCVACAHY KDPPFCVARC PSGVKPDLSY MPIWKFPDEE GACQPCPINC THSCVDLDDK GCPAEQRASP LTSIISAVVG ILLVVVLGVV FGILIKRRQQ KIRKYTMRRL LQETELVEPL TPSGAMPNQA QMRILKETEL RKVKVLGSGA FGTVYKGIWI PDGENVKIPV AIKVLRENTS PKANKEILDE AYVMAGVGSP YVSRLLGICL TSTVQLVTQL MPYGCLLDHV RENRGRLGSQ DLLNWCMQIA KGMSYLEDVR LVHRDLAARN VLVKSPNHVK ITDFGLARLL DIDETEYHAD GGKVPIKWMA LESILRRRFT HQSDVWSYGV TVWELMTFGA KPYDGIPARE IPDLLEKGER LPQPPICTID VYMIMVKCWM IDSECRPRFR ELVSEFSRMA RDPQRFVVIQ NEDLGPASPL DSTFYRSLLE DDDMGDLVDA EEYLVPQQGF FCPDPAPGAG GMVHHRHRSS STRNM (SEQ ID NO:999).

[0256] In some cases, a cancer-associated peptide is a peptide of a BCMA polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following BCMA amino acid sequence:

[0257] MLQMAGQCSQ NEYFDSLLHA CIPCQLRCSS NTPPLTCQRY CNASVTNSVK GTNAILWTCL GLSLIISLAV FVLMFLLRKI SSEPLKDEFK NTGSGLLGMA NIDLEKSRTG DEIILPRGLE YTVEECTCED CIKSKPKVDS DHCFPLPAME EGATILVTTK TNDYCKSLPA ALSATEIEKS ISAR (SEQ ID NO:1000).

[0258] In some cases, a cancer-associated peptide is a peptide of a WT-1 polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following WT-1 amino acid sequence:

[0259] MDFLLLQDPA STCVPEPASQ HTLRSGPGCL QQPEQQGVRD PGGIWAKLGA AEASAERLQG RRSRGASGSE PQQMGSDVRD LNALLPAVPS LGGGGGCALP VSGAAQWAPV LDFAPPGASA YGSLGGPAPP PAPPPPPPPP PHSFIKQEPS WGGAEPHEEQ CLSAFTVHFS GQFTGTAGAC RYGPFGPPPP SQASSGQARM FPNAPYLPSC LESQPAIRNQ GYSTVTFDGT PSYGHTPSHH AAQFPNHSFK HEDPMGQQGS LGEQQYSVPP PVYGCHTPTD SCTGSQALLL RTPYSSDNLY QMTSQLECMT WNQMNLGATL KGHSTGYESD NHTTPILCGA QYRIHTHGVF RGIQDVRRVP GVAPTLVRSA SETSEKRPFM CAYPGCNKRY FKLSHLQMHS RKHTGEKPYQ CDFKDCERRF SRSDQLKRHQ RRHTGVKPFQ CKTCQRKFSR SDHLKTHTRT HTGEKPFSCR WPSCQKKFAR SDELVRHHNM HQRNMTKLQL AL (SEQ ID NO:1001).

[0260] Non-limiting examples of WT-1 peptides include RMFPNAPYL (SEQ ID NO:1002), CMTWNQMN (SEQ ID NO:1003), CYTWNQMNL (SEQ ID NO:1004), CMTWNQMNLGATLKG (SEQ ID NO:1005), WNQMNLGATLKGVAA (SEQ ID NO:1006), CMTWNYMNLGATLKG (SEQ ID NO:1007), WNYMNLGATLKGVAA (SEQ ID NO:1008), MTWNQMNLGATLKGV (SEQ ID NO:1009), TWNQMNLGATLKGVA (SEQ ID NO:1010), CMTWNLMNLGATLKG (SEQ ID NO:1011), MTWNLMNLGATLKGV (SEQ ID NO:1012), TWNLMNLGATLKGVA (SEQ ID NO:1013), WNLMNLGATLKGVAA (SEQ ID NO:1014), MNLGATLK (SEQ ID NO:1015), MTWNYMNLGATLKGV (SEQ ID NO:1016), TWNYMNLGATLKGVA (SEQ ID NO:1017), CMTWNQMNLGATLKGVA (SEQ ID NO:1018), CMTWNLMNLGATLKGVA (SEQ ID NO:1019), CMTWNYMNLGATLKGVA (SEQ ID NO:1020), GYLRNPTAC (SEQ ID NO:1021), GALRNPTAL (SEQ ID NO:1022), YALRNPTAC (SEQ ID NO:1023), GLLRNPTAC (SEQ ID NO:1024), RYRPHPGAL (SEQ ID NO:1025), YQRPHPGAL (SEQ ID NO:1026), RLRPHPGAL (SEQ ID NO:1027), RIRPHPGAL (SEQ ID NO:1028), QFPNHSFKHEDPMGQ (SEQ ID NO:1029), HSFKHEDPY (SEQ ID NO:1030), QFPNHSFKHEDPM (SEQ ID NO:1031), QFPNHSFKHEDPY (SEQ ID NO:1032), KRPFMCAYPGCNK (SEQ ID NO:1033), KRPFMCAYPGCYK (SEQ ID NO:1034), FMCAYPGCY (SEQ ID NO:1035), FMCAYPGCK (SEQ ID NO:1036), KRPFMCAYPGCNKRY (SEQ ID NO:1037), SEKRPFMCAYPGCNK (SEQ ID NO:1038), KRPFMCAYPGCYKRY (SEQ ID NO:1039), NLMNLGATL (SEQ ID NO:1040), and NYMNLGATL (SEQ ID NO:1041).

[0261] In some cases, a cancer-associated peptide is a peptide of a human papillomavirus (HPV) polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to an HPV polypeptide. An HPV peptide can be a peptide of an HPV E6 polypeptide or an HPV E7 polypeptide. The HPV epitope can be an epitope of HPV of any of a variety of genotypes, including,e.g., HPV16, HPV18, HPV31, HPV33, HPV35, HPV39, HPV45, HPV51, HPV52, HPV56, HPV58, HPV59, HPV68, HPV73, or HPV82. Non-limiting examples of HPV peptides include: E618-26 (KLPQLCTEL; SEQ ID NO:1042); E626-34 (LQTTIHDII; SEQ ID NO:1043); E649-57 (VYDFAFRDL; SEQ ID NO:1044); E652-60 (FAFRDLCIV; SEQ ID NO:1045); E675-83 (KFYSKISEY; SEQ ID NO:1046); E680-88 (ISEYRHYCY; SEQ ID NO:1047); E77-15 (TLHEYMLDL; SEQ ID NO:1048); E711-19 (YMLDLQPET; SEQ ID NO:1049); E744-52 (QAEPDRAHY; SEQ ID NO:1050); E749-57 (RAHYNIVTF (SEQ ID NO:1051); E761-69 (CDSTLRLCV; SEQ ID NO:1052); and E767-76 (LCVQSTHVDI; SEQ ID NO:1053); E782-90 (LLMGTLGIV; SEQ ID NO:1054); E786-93 (TLGIVCPI; SEQ ID NO:1055); and E792-93 (LLMGTLGIVCPI; SEQ ID NO:1056).

[0262] In some cases, a cancer-associated peptide is a peptide of a claudin polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following claudin-18 (isoform 2) (CLDN 18.2) amino acid sequence:

[0263] MAVTACQGLG FVVSLIGIAG IIAATCMDQW STQDLYNNPV TAVFNYQGLW RSCVRESSGF TECRGYFTLL GLPAMLQAVR ALMIVGIVLG AIGLLVSIFA LKCIRIGSME DSAKANMTLT SGIMFIVSGL CAIAGVSVFA NMLVTNFWMS TANMYTGMGG MVQTVQTRYT FGAALFVGWV AGGLTLIGGV MMCIACRGLA PEETNYKAVS YHASGHSVAY KPGGFKASTG FGSNTKNKKI YDGGARTEDE VQSYPSKHDY V (SEQ ID NO:1057). In some cases, a cancer- associated peptide is a peptide of a claudin polypeptide having the amino acid sequence TEDEVQSYPSKHDYV (SEQ ID NO:1058) (and having a length of about 15 amino acids) or EVQSYPSKHDYV (SEQ ID NO:1059) (and having a length of about 12 amino acids.

[0264] In some cases, a cancer-associated peptide is a peptide of a trophoblast cell-surface antigen-2 (Trop-2) polypeptide. Trop-2 (also known as epithelial glycoprotein-1, gastrointestinal tumor- associated antigen GA733-1, membrane component chromosome 1 surface marker-1, and tumor- associated calcium signal transducer-2) is a transmembrane glycoprotein that is upregulated in numerous cancer types, and is the protein product of the TACSTD2 gene. In some cases, a cancer-associated peptide is a peptide of a TROP-2 polypeptide having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following TROP-2 amino acid sequence: QDNCTCPTNK MTVCSPDGPG GRCQCRALGS GMAVDCSTLT SKCLLLKARM SAPKNARTLV RPSEHALVDN DGLYDPDCDP EGRFKARQCN QTSVCWCVNS VGVRRTDKGD LSLRCDELVR THHILIDLRH RPTAGAFNHS DLDAELRRLF RERYRLHPKF VAAVHYEQPT IQIELRQNTS QKAAGDVDIG DAAYYFERDI KGESLFQGRG GLDLRVRGEP LQVERTLIYY LDEIPPKFSM KRLTAGLIAV IVVVVVALVA GMAVLVITNR RKSGKYKKVE IKELGELRKE PSL (SEQ ID NO:1060).Antibodies

[0265] As noted above, in some cases, a TTP present in a TMMP is an antibody. In some cases, the TTP is an antibody that is specific for a cancer-associated antigen. In some cases, the TTP is an antibody specific for a peptide / HLA complex on the surface of a cancer cell, where the peptide can be a cancer-associated peptide (e.g., a peptide of a cancer-associated antigen).

[0266] Non-limiting examples of cancer-associated antigen-targeted antibodies that can be included in a TMMP include, but are not limited to, abituzumab (anti-CD51), LL1 (anti-CD74), LL2 or RFB4 (anti-CD22), veltuzumab (hA20, anti-CD20), ^ binutuzu (anti-CD20), ^ binutuzumab (GA101, anti-CD20), daratumumab (anti-CD38), lambrolizumab (anti-PD-1 receptor), nivolumab (anti-PD-1 receptor), ipilimumab (anti-CTLA-4), RS7 (anti-TROP-2), PAM4 or KC4 (both anti-mucin), MN-14 (anti-CEA), MN-15 or MN-3 (anti-CEACAM6), Mu-9 (anti-colon-specific antigen-p), Immu 31 (anti- alpha-fetoprotein), R1 (anti-IGF-1R), A19 (anti-CD19), TAG-72 (e.g., CC49), Tn, J591 or HuJ591 (anti- PSMA), AB-PG1-XG1-026 (anti-PSMA dimer), D2 / B (anti-PSMA), G250 (anti-carbonic anhydrase IX), L243 (anti-HLA-DR) alemtuzumab (anti-CD52), oportuzumab (anti-EpCAM), bevacizumab (anti- VEGF), cetuximab (anti-EGFR), gemtuzumab (anti-CD33), ibritumomab tiuxetan (anti-CD20); panitumumab (anti-EGFR); tositumomab (anti-CD20); PAM4 (also known as clivatuzumab; anti-mucin), trastuzumab (anti-HER2), pertuzumab (anti-HER2), polatuzumab (anti-CD79b), and anetumab (anti- mesothelin).

[0267] In some cases, the tumor-targeting polypeptide is a single-chain antibody. In some cases, the tumor-targeting polypeptide is a scFv. In some cases, the tumor-targeting polypeptide is a nanobody (also referred to as a single domain antibody (sdAb)). In some cases, the tumor-targeting polypeptide is a heavy chain nanobody. In some cases, the tumor-targeting polypeptide is a light chain nanobody.

[0268] VH and VL amino acid sequences of various tumor antigen-binding antibodies are known in the art, as are the light chain and heavy chain CDRs of such antibodies. See, e.g., Ling et al. (2018) Frontiers Immunol.9:469; WO 2005 / 012493; US 2019 / 0119375; US 2013 / 0066055. The following are non-limiting examples of tumor antigen-binding antibodies. Anti-Her2

[0269] In some cases, an anti-Her2 antibody comprises: a) a light chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0270] DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLY SGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:1061); and b) a heavy chain comprising anamino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0271] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPT NGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTL VTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSS GLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSV LTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLV KGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEAL HNHYTQKSLSLSPGK (SEQ ID NO:1062).

[0272] In some cases, an anti-Her2 antibody comprises a light chain variable region (VL) present in the light chain amino acid sequence provided above; and a heavy chain variable region (VH) present in the heavy chain amino acid sequence provided above. For example, an anti-Her2 antibody can comprise: a) a VL comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (SEQ ID NO:1063); and b) a VH comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (SEQ ID NO:1064). In some cases, an anti-Her2 antibody comprises, in order from N-terminus to C-terminus: a) a VH comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSS (SEQ ID NO:1065); b) a linker; and c) a VL comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGS RSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (SEQ ID NO:1066). Suitable linkers are described elsewhere herein and include, e.g., (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10).

[0273] In some cases, an anti-Her2 antibody comprises VL CDR1, VL CDR2, and VL CDR3 present in the light chain amino acid sequence provided above; and VH CDR1, CDR2, and CDR3 present in the heavy chain amino acid sequence provided above. In some cases, the VH and VL CDRs areas defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0274] For example, an anti-Her2 antibody can comprise a VL CDR1 having the amino acid sequence RASQDVNTAVA (SEQ ID NO:1067); a VL CDR2 having the amino acid sequence SASFLY (SEQ ID NO:1068); a VL CDR3 having the amino acid sequence QQHYTTPP (SEQ ID NO:1069); a VH CDR1 having the amino acid sequence GFNIKDTY (SEQ ID NO:1070); a VH CDR2 having the amino acid sequence IYPTNGYT (SEQ ID NO:1071); and a VH CDR3 having the amino acid sequence SRWGGDGFYAMDY (SEQ ID NO:1072).

[0275] In some cases, an anti-Her2 antibody is a scFv antibody. For example, an anti-Her2 scFv can comprise an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADS VKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSGGGG SGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASF LYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (SEQ ID NO:1073).

[0276] As another example, in some cases, an anti-Her2 antibody comprises: a) a light chain variable region (VL) comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0277] DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSASYRY TGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKVEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:1074); and b) a heavy chain variable region (VH) comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0278] EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNP NSGGSIYNQRFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARNLGPSFYFDYWGQGTLVT VSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPP KPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALH NHYTQKSLSLSPG (SEQ ID NO:1075).

[0279] In some cases, an anti-Her2 antibody comprises a VL present in the light chain amino acid sequence provided above; and a VH present in the heavy chain amino acid sequence provided above. For example, an anti-Her2 antibody can comprise: a) a VL comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: DIQMTQSPSSLSASVGDRVTITCKASQDVSIGVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQQYYIYPYTFGQGTKVEIK (SEQ ID NO:1076); and b) a VH comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: EVQLVESGGGLVQPGGSLRLSCAASGFTFTDYTMDWVRQAPGKGLEWVADVNPNSGGSIYNQ RFKGRFTLSVDRSKNTLYLQMNSLRAEDTAVYYCARNLGPSFYFDYWGQGTLVTVSS (SEQ ID NO:1077).

[0280] In some cases, an anti-Her2 antibody comprises VL CDR1, VL CDR2, and VL CDR3 present in the light chain amino acid sequence provided above; and VH CDR1, CDR2, and CDR3 present in the heavy chain amino acid sequence provided above. In some cases, the VHand VLCDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VHand VLCDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0281] For example, an anti-HER2 antibody can comprise a VL CDR1 having the amino acid sequence KASQDVSIGVA (SEQ ID NO:1078); a VL CDR2 having the amino acid sequence SASYRY (SEQ ID NO:1079); a VL CDR3 having the amino acid sequence QQYYIYPY (SEQ ID NO:1080); a VH CDR1 having the amino acid sequence GFTFTDYTMD (SEQ ID NO:1081); a VH CDR2 having the amino acid sequence ADVNPNSGGSIYNQRFKG (SEQ ID NO:1082); and a VH CDR3 having the amino acid sequence ARNLGPSFYFDY (SEQ ID NO:1083).

[0282] In some cases, an anti-Her2 antibody is a scFv. For example, in some cases, an anti-Her2 scFv comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0283] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPT NGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTL VTVSSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKA PKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIK (SEQ ID NO:1073). Anti-CD19

[0284] Anti-CD19 antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-CD19 antibody can be used in a TMMP. See e.g., WO 2005 / 012493.

[0285] In some cases, an anti-CD19 antibody includes a VL CDR1 comprising the amino acid sequence KASQSVDYDGDSYLN (SEQ ID NO:1084); a VL CDR2 comprising the amino acid sequence DASNLVS (SEQ ID NO:1085); and a VL CDR3 comprising the amino acid sequence QQSTEDPWT (SEQ ID NO:1086). In some cases, an anti-CD19 antibody includes a VH CDR1 comprising the amino acid sequence SYWMN (SEQ ID NO:1087); a VH CDR2 comprising the amino acid sequence QIWPGDGDTNYNGKFKG (SEQ ID NO:1088); and a VH CDR3 comprising the amino acid sequence RETTTVGRYYYAMDY (SEQ ID NO:1089). In some cases, an anti-CD19 antibody includes a VL CDR1 comprising the amino acid sequence KASQSVDYDGDSYLN (SEQ ID NO:1084); a VL CDR2 comprising the amino acid sequence DASNLVS (SEQ ID NO:1085); a VL CDR3 comprising the amino acid sequence QQSTEDPWT (SEQ ID NO:1086); a VH CDR1 comprising the amino acid sequence SYWMN (SEQ ID NO:1087); a VH CDR2 comprising the amino acid sequence QIWPGDGDTNYNGKFKG (SEQ ID NO:1088); and a VH CDR3 comprising the amino acid sequence RETTTVGRYYYAMDY (SEQ ID NO:1089).

[0286] In some cases, an anti-CD19 antibody is a scFv. For example, in some cases, an anti- CD19 scFv comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRF SGSGSGTDFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGGSGGGGSGGGGSQVQ LQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFK GKATLTADESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAMDYWGQGTTVTVS (SEQ ID NO:1090). Anti-mesothelin

[0287] Anti-mesothelin antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-mesothelin antibody can be used in a TMMP. See, e.g., U.S.2019 / 0000944; WO 2009 / 045957; WO 2014 / 031476; USPN 8,460,660; US 2013 / 0066055; and WO 2009 / 068204. In some cases, the TTP is an anti-mesothelin scFv or an anti-mesothelin nanobody comprising VH and VL CDRs present in any one of the amino acid sequences set forth in FIG.22A-22H. In some cases, the TTP is an anti-mesothelin scFv comprising an amino acid sequence as set forth in any one of FIG.22A-22H.

[0288] In some cases, an anti-mesothelin antibody comprises: a) a light chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0289] DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNR PSGVSNRFSGSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKGDSSPVKAGVETTTPSKQSNNKYAASSYLSL TPEQWKSHRSYSCQVTHEGSTVEKTVAPTESS (SEQ ID NO:1091); and

[0290] b) a heavy chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0291] QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPG DSRTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLV TVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSG LYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFP PKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKG FYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK (SEQ ID NO:1092).

[0292] In some cases, an anti-mesothelin antibody comprises a VL present in the light chain amino acid sequence provided above; and a VH present in the heavy chain amino acid sequence provided above. For example, an anti-mesothelin antibody can comprise: a) a VL comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTK (SEQ ID NO:1093); and b) a VH comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSF QGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:1094).

[0293] In some cases, an anti-mesothelin antibody comprises VL CDR1, VL CDR2, and VL CDR3 present in the light chain amino acid sequence provided above; and VH CDR1, CDR2, and CDR3 present in the heavy chain amino acid sequence provided above. In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0294] For example, an anti-mesothelin antibody can comprise a VL CDR1 having the amino acid sequence TGTSSDIGGYNSVS (SEQ ID NO:1095); a VL CDR2 having the amino acid sequence LMIYGVNNRPS (SEQ ID NO:1096); a VL CDR3 having the amino acid sequence SSYDIESATP (SEQ ID NO:1097); a VH CDR1 having the amino acid sequence GYSFTSYWIG (SEQ ID NO:1098); aVH CDR2 having the amino acid sequence WMGIIDPGDSRTRYSP (SEQ ID NO:1099); and a VH CDR3 having the amino acid sequence GQLYGGTYMDG (SEQ ID NO:1100).

[0295] An anti-mesothelin antibody can be a scFv. As one non-limiting example, an anti- mesothelin scFv can comprise the following amino acid sequence: QVQLQQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGRINPNSGGTNYA QKFQGRVTMTRDTSISTAYMELSRLRSEDTAVYYCARGRYYGMDVWGQGTMVTVSSGGGGS GGGGSGGGGSGGGGSEIVLTQSPATLSLSPGERATISCRASQSVSSNFAWYQQRPGQAPRLLIYD ASNRATGIPPRFSGSGSGTDFTLTISSLEPED FAAYYCHQRSNWLYTFGQGTKVDIK (SEQ ID NO:1101), where VH CDR1, CDR2, and CDR3 are underlined; and VL CDR1, CDR2, and CDR3 are bolded and underlined.

[0296] As one non-limiting example, an anti-mesothelin scFv can comprise the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNSGGTNY AQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLRRTVVTPRAYYGMDVWGQGTTV TVSSGGGGSGGGGSGGGGSGGGGSDIQLTQSPSTLSASVGDRVTITCQASQDISNSLNWYQQKA GKAPKLLIYDASTLETGVPSRFSGSGSGTDFSF TISSLQPEDIATYYCQQHDNLPLTFGQGTKVEIK (SEQ ID NO:1102), where VH CDR1, CDR2, and CDR3 are underlined; and VL CDR1, CDR2, and CDR3 are bolded and underlined.

[0297] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIK (SEQ ID NO:1103); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QMQLVESGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYV DSVKGRFTISRDNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSS (SEQ ID NO:1104). In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987). See, e.g., BMS6A5.

[0298] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIK (SEQ ID NO:1103); and b) a VHregion comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QMQLVESGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYV DSVKGRFTISRDNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSS (SEQ ID NO:1104).

[0299] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QMQLVESGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYV DSVKGRFTISRDNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSS (SEQ ID NO:1104); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIK (SEQ ID NO:1103). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0300] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIK (SEQ ID NO:1103); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QMQLVESGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYV DSVKGRFTISRDNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSS (SEQ ID NO:1104). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0301] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising the following amino acid sequence: DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIK (SEQ ID NO:1105); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVQLQQSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKF RGKATLTVDKSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSS (SEQ ID NO:1106). In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987). See, e.g., Amatuximab.

[0302] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIK (SEQ ID NO:1105); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKF RGKATLTVDKSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSS (SEQ ID NO:1106).

[0303] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIK (SEQ ID NO:1105); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKF RGKATLTVDKSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSS (SEQ ID NO:1106). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases,the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0304] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKF RGKATLTVDKSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSS (SEQ ID NO:1106); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIK (SEQ ID NO:1105). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0305] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising the following amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLG (SEQ ID NO:1107); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSF QGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:1094). In some cases, the VHand VLCDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VHand VLCDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0306] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLG (SEQ ID NO:1107); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSF QGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:1094).

[0307] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLG (SEQ ID NO:1107); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSF QGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:1094). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0308] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSF QGQVTISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:1094); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLG (SEQ ID NO:1107). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0309] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising thefollowing amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFSGSG SGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK (SEQ ID NO:1108); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYN QKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSS (SEQ ID NO:1109). In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987). See, e.g., RG7787.

[0310] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFSGSG SGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK (SEQ ID NO:1108); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYN QKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSS (SEQ ID NO:1109).

[0311] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFSGSG SGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK (SEQ ID NO:1108); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYN QKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSS (SEQ ID NO:1109). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0312] In some cases, an anti-mesothelin antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYN QKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSS (SEQ ID NO:1109); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFSGSG SGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK (SEQ ID NO:1108). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0313] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: QMQLVESGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYVDSVK GRFTISRDNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSSGGGGSGGG GSGGGGSEIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATG IPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIK (SEQ ID NO:888), where the VH sequence is italicized, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is underlined.

[0314] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: EIVLTQSPGTLSLSPGERATLSCRASQSVSSSYLAWYQQKPGQAPRLLIYGASSRATGIPDRFSGS GSGTDFTLTISRLEPEDFAVYYCQQYGSSPIFTFGPGTKVDIKGGGGSGGGGSGGGGSQMQLVE SGGGVVQPGRSLRLSCTASGFTFSNNGMHWVRQAPGKGLEWVAVIWFDGMNKFYVDSVKGRFTISR DNSKNTLYLEMNSLRAEDTAIYYCAREGDGSGIYYYYGMDVWGQGTTVTVSS (SEQ ID NO:889), where the VL sequence is underlined, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is italicized.

[0315] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: QVQLQQSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKFRGKA TLTVDKSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSSGGGGSGGGGSGGGGS DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIK (SEQ ID NO:890), where the VHsequence is italicized, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is underlined.

[0316] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: DIELTQSPAIMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPGRFSGS GSGNSYSLTISSVEAEDDATYYCQQWSKHPLTFGSGTKVEIKGGGGSGGGGSGGGGSQVQLQ QSGPELEKPGASVKISCKASGYSFTGYTMNWVKQSHGKSLEWIGLITPYNGASSYNQKFRGKATLTVD KSSSTAYMDLLSLTSEDSAVYFCARGGYDGRGFDYWGSGTPVTVSS (SEQ ID NO:891), where the VL sequence is underlined, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is italicized.

[0317] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: QVELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSFQGQV TISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSSGGGGSGGGGSGGG GSDIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNR FSGSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLG (SEQ ID NO:892), where the VH sequence is italicized, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is underlined.

[0318] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: DIALTQPASVSGSPGQSITISCTGTSSDIGGYNSVSWYQQHPGKAPKLMIYGVNNRPSGVSNRFS GSKSGNTASLTISGLQAEDEADYYCSSYDIESATPVFGGGTKLTVLGGGGGSGGGGSGGGGSQ VELVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQAPGKGLEWMGIIDPGDSRTRYSPSFQGQVT ISADKSISTAYLQWSSLKASDTAMYYCARGQLYGGTYMDGWGQGTLVTVSS (SEQ ID NO:893), where the VL sequence is underlined, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is italicized.

[0319] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYNQKFRG KATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSSGGGGSGGGGSGG GGSDIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFS GSGSGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIK (SEQ ID NO:894), where the VH sequence is italicized, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is underlined.

[0320] In some cases, an anti-mesothelin scFv comprises the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASSSVSYMHWYQQKSGKAPKLLIYDTSKLASGVPSRFSGSG SGTDFTLTISSLQPEDFATYYCQQWSKHPLTFGQGTKLEIKGGGGSGGGGSGGGGSQVQLVQS GAEVKKPGASVKVSCKASGYSFTGYTMNWVRQAPGQGLEWMGLITPYNGASSYNQKFRGKATMTVDTSTSTVYMELSSLRSEDTAVYYCARGGYDGRGFDYWGQGTLVTVSS (SEQ ID NO:895), where the VL sequence is underlined, the (GGGGS)3 (SEQ ID NO:875) linker is bolded and underlined, and the VL sequence is italicized. Anti-PSMA

[0321] Prostate-specific membrane antigen (PSMA) (also known as folate hydrolase 1 (FOLH1); membrane glutamate carboxypeptidase, and N-Acetylated-Alpha-Linked Acidic Dipeptidase 1) that is up-regulated in cancerous cells in the prostate and is used as a diagnostic and prognostic indicator of prostate cancer.

[0322] Anti-PSMA antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-PSMA antibody can be used in a TMMP. See, e.g., U.S. Patent No.10,179,819 and U.S. Patent Publication No.2021 / 0277141. Anti-CD22

[0323] CD22 (also known as B-Lymphocyte Cell Adhesion Molecule, Sialic Acid-Binding Ig- Like Lectin 2, or SIGLEC2) is a sialic acid-binding adhesion molecule largely restricted to the B cell lineage and expressed on most B-lineage malignancies.

[0324] Anti-CD22 antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-CD22 antibody can be used in a TMMP. See, e.g., Xiao et al. (2009) Mabs 1:297 (describing the fully human anti-CD22 m971 scFv); and U.S. Patent Publication No.2020 / 0147134. Examples of anti-CD22 antibodies include epratuzumab and inotuzumab. See, e.g., Lenoard et al. (2007) Oncogene 26:3704 and U.S. Patent No.5,789,554 (describing epratuzumab); and DiJoseph et al. (2007) Leukemia 21:2240 (describing inotuzumab).

[0325] For example, an anti-CD22 antibody can comprise: i) a heavy chain variable region (VH) CDR1 having the amino acid sequence: GDSVSSNSAA (SEQ ID NO:1110); ii) a VH CDR2 having the amino acid sequence: TYYRSKWYN (SEQ ID NO:1111); iii) a VH CDR3 having the amino acid sequence: AREVTGDLEDAFDI (SEQ ID NO:1112); iv) a light chain variable region (VL) CDR1 having the amino acid sequence: QTIWSY (SEQ ID NO:1113); v) a VL CDR2 having the amino acid sequence: AAS (Ala-Ala-Ser); and vi) a VL CDR3 having the amino acid sequence: QQSYSIPQT (SEQ ID NO:1114). Anti-TROP-2

[0326] Trophoblast cell surface antigen 2 (Trop-2) (also known as epithelial glycoprotein-1, gastrointestinal tumor-associated antigen GA733-1, membrane component chromosome 1 surface marker-1, and tumor-associated calcium signal transducer-2) is a transmembrane glycoprotein that is upregulated in numerous cancer types, and is the protein product of the TACSTD2 gene.

[0327] In some cases, the TTP is an anti-TROP-2 scFv or an anti-TROP-2 nanobody comprising VH and VL CDRs present in any one of the amino acid sequences set forth in FIG.23A-23D. In some cases, the TTP is an anti-TROP-2 scFv comprising an amino acid sequence as set forth in any one of FIG.23A-23D.

[0328] Anti-TROP-2 antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-TROP-2 antibody can be used in a TMMP. See, e.g., U.S. Patent No.7,238,785). In some cases, an anti-TROP-2 antibody comprises: i) light chain CDR sequences CDR1 (KASQDVSIAVA; SEQ ID NO:1115); CDR2 (SASYRYT; SEQ ID NO:1116); and CDR3 (QQHYITPLT; SEQ ID NO:1117); and ii) heavy chain CDR sequences CDR1 (NYGMN; SEQ ID NO:1118); CDR2 (WINTYTGEPTYTDDFKG; SEQ ID NO:1119); and CDR3 (GGFGSSYWYFDV; SEQ ID NO:1120).

[0329] In some cases, an anti-TROP-2 antibody comprises: i) heavy chain CDR sequences CDR1 (TAGMQ; SEQ ID NO:1121); CDR2 (WINTHSGVPKYAEDFKG (SEQ ID NO:1122); and CDR3 (SGFGSSYWYFDV; SEQ ID NO:1123); and ii) light chain CDR sequences CDR1 (KASQDVSTAVA; SEQ ID NO:1124); CDR2 (SASYRYT; SEQ ID NO:1116); and CDR3 (QQHYITPLT; SEQ ID NO:1117).

[0330] In some cases, an anti-TROP2 antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising the following amino acid sequence: DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGS GSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK (SEQ ID NO:1125); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYT DDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS (SEQ ID NO:1126). In some cases, the VHand VLCDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0331] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGS GSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK (SEQ ID NO:1125); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%,or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYT DDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS (SEQ ID NO:1126).

[0332] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGS GSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK (SEQ ID NO:1125); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYT DDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS (SEQ ID NO:1126). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0333] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYT DDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSS (SEQ ID NO:1126); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQLTQSPSSLSASVGDRVSITCKASQDVSIAVAWYQQKPGKAPKLLIYSASYRYTGVPDRFSGS GSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGAGTKVEIK (SEQ ID NO:1125). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0334] In some cases, an anti-TROP2 antibody suitable for inclusion in a TMMP comprises: a) VL CDR1, VL CDR2, and VL CDR3 present in a light chain variable region (VL) comprising thefollowing amino acid sequence: DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSG SGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQGTKLEIK (SEQ ID NO:1127); and b) VH CDR1, CDR2, and CDR3 present in a heavy chain variable region (VH) comprising the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQGLEWMGWINTHSGVPKYA EDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCARSGFGSSYWYFDVWGQGTLVTVSS (SEQ ID NO:1128). In some cases, the VH and VL CDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VH and VL CDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0335] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP comprises: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSG SGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQGTKLEIK (SEQ ID NO:1127); and b) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQGLEWMGWINTHSGVPKYA EDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCARSGFGSSYWYFDVWGQGTLVTVSS (SEQ ID NO:1128).

[0336] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSG SGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQGTKLEIK (SEQ ID NO:1127); b) a peptide linker; and c) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQGLEWMGWINTHSGVPKYA EDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCARSGFGSSYWYFDVWGQGTLVTVSS (SEQ ID NO:1128). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids.

[0337] In some cases, an anti-TROP-2 antibody suitable for inclusion in a TMMP is a scFv comprising, in order from N-terminus to C-terminus: a) a VH region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGASVKVSCKASGYTFTTAGMQWVRQAPGQGLEWMGWINTHSGVPKYA EDFKGRVTISADTSTSTAYLQLSSLKSEDTAVYYCARSGFGSSYWYFDVWGQGTLVTVSS (SEQ ID NO:1128); b) a peptide linker; and c) a VL region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCKASQDVSTAVAWYQQKPGKAPKLLIYSASYRYTGVPSRFSG SGSGTDFTLTISSLQPEDFAVYYCQQHYITPLTFGQGTKLEIK (SEQ ID NO:1127). In some cases, the peptide linker comprises the amino acid sequence (GGGGS)n (SEQ ID NO:934), where n is an integer from 1 to 10 (e.g., where n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10). In some cases, the peptide linker comprises the amino acid sequence GGGGSGGGGSGGGGS (SEQ ID NO:875) and has a length of 15 amino acids. Anti-BCMA

[0338] Anti-BCMA (B-cell maturation antigen) antibodies are known in the art; and the VH and VL, or the VH and VL CDRs, of any anti-BCMA antibody can be used in a TMMP. See, e.g., WO 2014 / 089335; US 2019 / 0153061; and WO 2017 / 093942.

[0339] In some cases, an anti-BCMA antibody comprises: a) a light chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence:

[0340] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIFNYHQRP SGVPDRFSGSKSGSSASLAISGLQSEDEADYYCAAWDDSLNGWVFGGGTKLTVLGQPKAAPSV TLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPDSKQSNNKYAASSYL SLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO:1129); and

[0341] b) a heavy chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: EVQLVESGGGLVKPGGSLRLSCAASGFTFGDYALSWFRQAPGKGLEWVGVSRSKAYGGTTDY AASVKGRFTISRDDSKSTAYLQMNSLKTEDTAVYYCASSGYSSGWTPFDYWGQGTLVTVSSAS TKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSS VVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKD TLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQ DWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQ KSLSLSPGK (SEQ ID NO:1130).

[0342] In some cases, an anti-BCMA antibody comprises a VL present in the light chain amino acid sequence provided above; and a VH present in the heavy chain amino acid sequence provided above. For example, an anti-BCMA antibody can comprise: a) a VL comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence:

[0343] QSVLTQPPSASGTPGQRVTISCSGSSSNIGSNTVNWYQQLPGTAPKLLIFNYHQRP SGVPDRFSGSKSGSSASLAISGLQSEDEADYYCAAWDDSLNGWVFGGGTKLTVLG (SEQ ID NO:1131); and b) a VH comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence:

[0344] EVQLVESGGGLVKPGGSLRLSCAASGFTFGDYALSWFRQAPGKGLEWVGVSRS KAYGGTTDYAASVKGRFTISRDDSKSTAYLQMNSLKTEDTAVYYCASSGYSSGWTPFDYWGQ GTLVTVSSASTKGPSV (SEQ ID NO:1132).

[0345] In some cases, an anti-BCMA antibody comprises VL CDR1, VL CDR2, and VL CDR3 present in the light chain amino acid sequence provided above; and VH CDR1, CDR2, and CDR3 present in the heavy chain amino acid sequence provided above. In some cases, the VHand VLCDRs are as defined by Kabat (see, e.g., Table 1, above; and Kabat 1991). In some cases, the VHand VLCDRs are as defined by Chothia (see, e.g., Table 1, above; and Chothia 1987).

[0346] For example, an anti-BCMA antibody can comprise a VL CDR1 having the amino acid sequence SSNIGSNT (SEQ ID NO:1133), a VL CDR2 having the amino acid sequence NYH, a VL CDR3 having the amino acid sequence AAWDDSLNGWV (SEQ ID NO:1134)), a VH CDR1 having the amino acid sequence GFTFGDYA (SEQ ID NO:1135), a VH CDR2 having the amino acid sequence SRSKAYGGTT (SEQ ID NO:1136), and a VH CDR3 having the amino acid sequence ASSGYSSGWTPFDY (SEQ ID NO:1137).

[0347] An anti-BCMA antibody can be a scFv. As one non-limiting example, an anti-BCMA scFv can comprise the following amino acid sequence: QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYY NQKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSSGG GGSGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKL LIYYTSNLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKR (SEQ ID NO:1138).

[0348] As another example, an anti-BCMA scFv can comprise the following amino acid sequence:DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKRGGGGSGGGGSGGGGSGGGGSQ VQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYN QKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYNGYDVLDNWGQGTLVTVSS (SEQ ID NO:1139).

[0349] In some cases, an anti-BCMA antibody can comprise a VL CDR1 having the amino acid sequence SASQDISNYLN (SEQ ID NO:1140); a VL CDR2 having the amino acid sequence YTSNLHS (SEQ ID NO:1141); a VL CDR3 having the amino acid sequence QQYRKLPWT (SEQ ID NO:1142); a VH CDR1 having the amino acid sequence NYWMH (SEQ ID NO:1143); a VH CDR2 having the amino acid sequence ATYRGHSDTYYNQKFKG (SEQ ID NO:1144); and a VH CDR3 having the amino acid sequence GAIYNGYDVLDN (SEQ ID NO:1145).

[0350] In some cases, an anti-BCMA antibody comprises: a) a light chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKR (SEQ ID NO:1146).

[0351] In some cases, an anti-BCMA antibody comprises: a) a heavy chain comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following amino acid sequence: QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYY NQKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSS (SEQ ID NO:1147).

[0352] In some cases, an anti-BCMA antibody (e.g., an antibody referred to in the literature as belantamab) comprises a light chain comprising the amino acid sequence: DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKLLIYYTSNLHSGVPSRFSGS GSGTDFTLTISSLQPEDFATYYCQQYRKLPWTFGQGTKLEIKR (SEQ ID NO:1146); and a heavy chain comprising the amino acid sequence: QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYY NQKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSS (SEQ ID NO:1147).

[0353] In some cases, the anti-BCMA antibody has a cancer chemotherapeutic agent linked to the antibody. For example, in some cases, the anti-BCMA antibody is GSK2857916 (belantamab- mafodotin), where monomethyl auristatin F (MMAF) is linked via a maleimidocaproyl linker to the anti- BCMA antibody belantamab.Anti-MUC1

[0354] In some cases, a TTP present in a TMMP is an antibody specific for MUC1. For example, a TTP can be specific for a MUC1 polypeptide present on a cancer cell. In some cases, the TTP is specific for the cleaved form of MUC1; see, e.g., Fessler et al. (2009) Breast Cancer Res. Treat. 118:113. In some cases, the TTP is an antibody specific for a glycosylated MUC1 peptide; see, e.g., Naito et al. (2017) ACS Omega 2:7493; and US 10,017,580.

[0355] As one non-limiting example, a TTP can be a single-chain Fv specific for MUC1. See, e.g., Singh et al. (2007) Mol. Cancer Ther.6:562; Thie et al. (2011) PloSOne 6:e15921; Imai et al. (2004) Leukemia 18:676; Posey et al. (2016) Immunity 44:1444; EP3130607; EP3164418; WO 2002 / 044217; and US 2018 / 0112007. In some cases, a TTP is a scFv specific for the MUC1 peptide VTSAPDTRPAPGSTAPPAHG (SEQ ID NO:1148). In some cases, a TTP is a scFv specific for the MUC1 peptide SNIKFRPGSVVVQLTLAFREGTINVHDVETQFNQYKTEAASRY (SEQ ID NO:1149). In some cases, a TTP is a scFv specific for the MUC1 peptide SVVVQLTLAFREGTINVHDVETQFNQYKTEAASRY (SEQ ID NO:1150). In some cases, a TTP is a scFv specific for the MUC1 peptide LAFREGTINVHDVETQFNQY (SEQ ID NO:1151). In some cases, a TTP is a scFv specific for the MUC1 peptide SNIKFRPGSVVVQLTLAAFREGTIN (SEQ ID NO:1152).

[0356] As an example, an anti-MUC1 antibody can comprise: a VH CDR1 having the amino acid sequence RYGMS (SEQ ID NO:1153); a VH CDR2 having the amino acid sequence TISGGGTYIYYPDSVKG (SEQ ID NO:1154); a VH CDR3 having the amino acid sequence DNYGRNYDYGMDY (SEQ ID NO:1155); a VL CDR1 having the amino acid sequence SATSSVSYIH (SEQ ID NO:1156); a VL CDR2 having the amino acid sequence STSNLAS (SEQ ID NO:1157); and a VL CDR3 having the amino acid sequence QQRSSSPFT (SEQ ID NO:1158). See, e.g., US 2018 / 0112007.

[0357] As another example, an anti-MUC1 antibody can comprise a VH CDR1 having the amino acid sequence GYAMS (SEQ ID NO:1159); a VH CDR2 having the amino acid sequence TISSGGTYIYYPDSVKG (SEQ ID NO:1160); a VH CDR3 having the amino acid sequence LGGDNYYEYFDV (SEQ ID NO:1161); a VL CDR1 having the amino acid sequence RASKSVSTSGYSYMH (SEQ ID NO:1162); a VL CDR2 having the amino acid sequence LASNLES (SEQ ID NO:1163); and a VL CDR3 having the amino acid sequence QHSRELPFT (SEQ ID NO:1164). See, e.g., US 2018 / 0112007.

[0358] As another example, an anti-MUC1 antibody can comprise a VH CDR1 having the amino acid sequence DYAMN (SEQ ID NO:1165); a VH CDR2 having the amino acid sequence VISTFSGNINFNQKFKG (SEQ ID NO:1166); a VH CDR3 having the amino acid sequenceSDYYGPYFDY (SEQ ID NO:1167); a VL CDR1 having the amino acid sequence RSSQTIVHSNGNTYLE (SEQ ID NO:1168); a VL CDR2 having the amino acid sequence KVSNRFS (SEQ ID NO:1169); and a VL CDR3 having the amino acid sequence (FQGSHVPFT (SEQ ID NO:1170). See, e.g., US 2018 / 0112007.

[0359] As another example, an anti-MUC1 antibody can comprise a VH CDR1 having the amino acid sequence GYAMS (SEQ ID NO:1159); a VH CDR2 having the amino acid sequence TISSGGTYIYYPDSVKG (SEQ ID NO:1160); a VH CDR3 having the amino acid sequence LGGDNYYEY (SEQ ID NO:1171); a VL CDR1 having the amino acid sequence TASKSVSTSGYSYMH (SEQ ID NO:1172); a VL CDR2 having the amino acid sequence LVSNLES (SEQ ID NO:1173); and a VL CDR3 having the amino acid sequence QHIRELTRSE (SEQ ID NO:1174). See, e.g., US 2018 / 0112007. Anti-MUC16

[0360] In some cases, a TTP present in a TMMP is an antibody specific for MUC16 (also known as CA125). See, e.g., Yin et al. (2002) Int. J. Cancer 98:737. For example, a TTP can be specific for a MUC16 polypeptide present on a cancer cell. See, e.g., US 2018 / 0118848; and US 2018 / 0112008. In some cases, a MUC16-specific TTP is a scFv. In some cases, a MUC16-specific TTP is a nanobody.

[0361] As one example, an anti-MUC16 antibody can comprise a VH CDR1 having the amino acid sequence GFTFSNYY (SEQ ID NO:1175); a VH CDR2 having the amino acid sequence ISGRGSTI (SEQ ID NO:1176); a VH CDR3 having the amino acid sequence VKDRGGYSPY (SEQ ID NO:1177); a VL CDR1 having the amino acid sequence QSISTY (SEQ ID NO:1178); a VL CDR2 having the amino acid sequence TAS; and a VL CDR3 having the amino acid sequence QQSYSTPPIT (SEQ ID NO:1179). See, e.g., US 2018 / 0118848. Anti-Claudin-18.2

[0362] In some cases, a TTP present in a TMMP is an antibody specific for claudin-18 isoform 2 (“claudin-18.2”). See, e.g., WO 2013 / 167259. In some cases, a claudin-18.2-specific TTP is a scFv. In some cases, a claudin-18.2-specific TTP is a nanobody. In some cases, a TPP present in a TMMP is an antibody specific for TEDEVQSYPSKHDYV (SEQ ID NO:1058) or EVQSYPSKHDYV (SEQ ID NO:1059).

[0363] As one example, an anti-claudin-18.2 antibody can comprise a VH CDR1 having the amino acid sequence GYTFTDYS (SEQ ID NO:1180); a VH CDR2 having the amino acid sequence INTETGVP (SEQ ID NO:1181); a VH CDR3 having the amino acid sequence ARRTGFDY (SEQ ID NO:1182); a VL CDR1 having the amino acid sequence KNLLHSDGITY (SEQ ID NO:1183); a VL CDR2 having the amino acid sequence RVS; and a VL CDR3 having the amino acid sequence VQVLELPFT (SEQ ID NO:1184).

[0364] As another example, an anti-claudin-18.2 antibody can comprise a VH CDR1 having the amino acid sequence GFTFSSYA (SEQ ID NO:1185); a VH CDR2 having the amino acid sequence ISDGGSYS (SEQ ID NO:1186); a VH CDR3 having the amino acid sequence ARDSYYDNSYVRDY (SEQ ID NO:1187); a VL CDR1 having the amino acid sequence QDINTF (SEQ ID NO:1188); a VL CDR2 having the amino acid sequence RTN; and a VL CDR3 having the amino acid sequence LQYDEFPLT (SEQ ID NO:1189). mAb FUSIONS

[0365] In embodiments, a TMMP can comprise a heterodimer, wherein: a) the first polypeptide of the heterodimer comprises i) a single-chain polypeptide comprising a peptide epitope (e.g., a viral epitope); a β2M polypeptide; and an MHC class I heavy chain polypeptide, and ii) an antibody heavy chain polypeptide; and wherein b) the second polypeptide of the heterodimer comprises an antibody light chain polypeptide, and wherein either the first polypeptide or the second polypeptide comprises one or more immunomodulatory polypeptides, e.g., on the N-terminus of the antibody light chain polypeptide, on the C-terminus of the antibody light chain polypeptide, or on the C-terminus of the antibody heavy chain polypeptide. A TMMP can thus comprise a heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope (e.g., a viral epitope); ii) a β2M polypeptide; iii) an MHC class I heavy chain polypeptide; and iv) an antibody heavy chain polypeptide (e.g., an antibody heavy chain comprising, in order from N-terminus to C-terminus: i) a VH polypeptide; ii) an Ig CH1 polypeptide; and iii) an IgG Fc polypeptide); and b) a second polypeptide comprising: i) one or more immunomodulatory polypeptides; and ii) an antibody light chain polypeptide (e.g., an antibody light chain comprising, in order from N-terminus to C-terminus: i) a VL polypeptide; and an Ig kappa light chain constant region polypeptide). A TMMP can comprise a heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope (e.g., a viral epitope); ii) a β2M polypeptide; iii) an MHC class I heavy chain polypeptide; iv) an antibody heavy chain polypeptide (e.g., an antibody heavy chain comprising, in order from N-terminus to C-terminus: i) a VH polypeptide; ii) an Ig CH1 polypeptide; and iii) an IgG Fc polypeptide); and v) one or more immunomodulatory polypeptides; and b) a second polypeptide comprising an antibody light chain polypeptide (e.g., an antibody light chain comprising, in order from N-terminus to C-terminus: i) a VL polypeptide; and an Ig kappa light chain constant region polypeptide). A TMMP can comprise a heterodimer comprising: a) a first polypeptide comprising: i) a peptide epitope (e.g., a viral epitope); ii) a β2M polypeptide; iii) an MHC class I heavy chain polypeptide; and iv) an antibody heavy chain polypeptide (e.g., an antibody heavy chain comprising, in order from N-terminus to C-terminus: i) a VH polypeptide; ii) an Ig CH1 polypeptide; and iii) an IgG Fc polypeptide); and b) a second polypeptide comprising: i) an antibody light chain polypeptide (e.g., an antibody light chain comprising, in order from N-terminus to C-terminus: i) a VL polypeptide; and an Ig kappa light chain constant region polypeptide); and ii) one or more immunomodulatory polypeptides. Such heterodimerscan dimerize (e.g., form homodimers of two of the heterodimers by the formation of disulfide bonds that form between the antibody heavy chain polypeptides of each heterodimer). Such TMMPs are referred to herein as “monoclonal antibody fusions” or “mAb fusions.” The VH polypeptide and the VL polypeptide are on separate polypeptides of the heterodimer; together, the VH polypeptide and the VL polypeptide bind to a cancer epitope and therefore constitute a TTP.

[0366] The following are non-limiting examples of mAb fusions.

[0367] As one example, a TMMP can comprise: a) a first polypeptide comprising the amino acid sequence depicted in FIG.24A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.24B. As an example, the SARS-CoV-2 peptide can have the amino acid sequence YLQPRTFLL (SEQ ID NO:218).

[0368] As another example, a TMMP can comprise: a) a first polypeptide comprising the amino acid sequence depicted in FIG.25A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.25B. As an example, the SARS-CoV-2 peptide can have the amino acid sequence YLQPRTFLL (SEQ ID NO:218).

[0369] As another example, a TMMP can comprise: a) a first polypeptide comprising the amino acid sequence depicted in FIG.26A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.26B. As an example, the SARS-CoV-2 peptide can have the amino acid sequence YLQPRTFLL (SEQ ID NO:218). Single-chain T-cell Receptors

[0370] As noted above, in some cases, a TTP present in a TMMP is a scTCR. A TTP can be a scTCR specific for a peptide / HLA complex on the surface of a cancer cell, where the peptide can be a cancer-associated peptide (e.g., a peptide of a cancer-associated antigen). Amino acid sequences of scTCRs specific for cancer-associated peptides bound to an HLA complex are known in the art. See, e.g., US 2019 / 0135914; US 2019 / 0062398; and US 2018 / 0371049.

[0371] A scTCR includes an alpha chain variable region (Vα) and a beta chain variable region (Vβ) covalently linked through a suitable peptide linker sequence. For example, the Vα can be covalently linked to the Vβ through a suitable peptide linker (L) sequence fused to the C-terminus of the Vα and the N-terminus of the Vβ. An scTCR can have the structure Vα-L-Vβ. An scTCR can have the structure Vβ- L-Vα. An scTCR can also comprise a constant domain (also referred to as constant region). In some cases, an scTCR comprises, in order from N-terminus to C-terminus: i) a TCR α chain variable domain polypeptide; ii) a peptide linker; iii) a TCR β chain variable domain polypeptide; and iv) a TCR β chain constant region extracellular domain polypeptide. In some cases, an scTCR comprises, in order from N- terminus to C-terminus: i) a TCR β chain variable domain polypeptide; ii) a peptide linker; iii) a TCR αchain variable domain polypeptide; and iv) a TCR α chain constant region extracellular domain polypeptide.

[0372] Amino acid sequences of scTCRs specific for peptide / HLA complexes, where the peptide is a cancer-associated peptide, are known in the art. See, e.g., US 2019 / 0135914; US 2019 / 0062398; US 2018 / 0371049; US 2019 / 0144563; and US 2019 / 0119350.

[0373] For example, a scTCR can be specific for an NY-ESO epitope such as an SLLMWITQC (SEQ ID NO:1190) peptide bound to an HLA complex comprising an HLA-A*0201 heavy chain and a β2M polypeptide. As an example, such an scTCR can comprise: i) a TCR α chain variable region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: MQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIQSSQREQTSGRLNAS LDKSSGRSTLYIAASQPGDSATYLCAVRPTSGGSYIPTFGRGTSLIVHPY (SEQ ID NO:1191), where amino acid 20 can be V or A; amino acid 51 can be Q, P, S, T, or M; amino acid 52 can be S, P, F, or G, amino acid 53 can be S, W, H, or T; amino acid 94 can be P, H, or A; amino acid 95 can be T, L, M, A, Q, Y, E, I, F, V, N, G, S, D, or R; amino acid 96 can be S, L, T, Y, I, Q, V, E, A, W, R, G, H, D, or K; amino acid 97 can be G, D, N, V, S, T, or A; amino acid 98 can be G, P, H, S, T, W, or A; amino acid 99 can be S, T, Y, D, H, V, N, E, G, Q, K, A, I, or R; amino acid 100 can be Y, F, M, or D; amino acid 101 can be I, P, T, or M; and amino acid 103 can be T or A; and ii) a TCR β chain variable region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: MGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVGAGITDQGEVPN GYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVL (SEQ ID NO:1192), where amino acid 18 can be M or V; amino acid 50 can be G, V, or I; amino acid 52 can be G or Q; amino acid 53 can be I, T, or M; amino acid 55 can be D or R; amino acid 56 can be Q or R; amino acid 70 can be T or I; amino acid 94 can be Y, N, or F; amino acid 95can be V or L; and amino acid 97 can be N, G, or D. For example, in some cases, a scTCR can comprise: i) a TCR α chain variable region comprising the amino acid sequence: MQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLTSLLLIMSHQREQTSGRLNA SLDKSSGRSTLYIAASQPGDSATYLCAVRPTSGGSYIPTFGRGTSLIVHPY (SEQ ID NO:1193); and a TCR β chain variable region comprising the amino acid sequence: MGVTQTPKFQVLKTGQSMTLQCAQDMNHEYMSWYRQDPGMGLRLIHYSVSAGITDQGEVPN GYNVSRSTTEDFPLRLLSAAPSQTSVYFCASSYVGNTGELFFGEGSRLTVL (SEQ ID NO:1194).

[0374] As another example, a scTCR can be specific for an HPV epitope (e.g., an HPV peptide of the amino acid sequence YIIFVYIPL (HPV 16 E563-71; SEQ ID NO:1195), KLPQLCTEL (HPV 16 E611-19; SEQ ID NO:1042), TIHEIILECV (HPV 16 E6; SEQ ID NO:1196), YMLDLQPET (HPV 16E711-19; SEQ ID NO:1049), TLGIVCPI (HPV 16 E786-93) (SEQ ID NO:1055), KCIDFYSRI (HPV 18 E667-75; SEQ ID NO:1197), or FQQLFLNTL (HPV 18 E786-94; SEQ ID NO:1198)) bound to an HLA complex comprising an HL heavy chain and a β2M polypeptide. As an example, such an scTCR can comprise: i) a TCR α chain variable region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: METLLGLLILQLQLQWVSSKQEVTQIPAALSVPEGENLVLNCSFTDSAIYNLQWFRQDPGKGLT SLLLIQSSQREQTSGRLNASLDKSSGRSTLYIAASQPGDSATYLCAVRETSGSRLTFGEGTQLTVN PD(SEQ ID NO:1199); and ii) a TCR β chain variable region comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence: MGIRLLCRVAFCFLAVGLVDVKVTQSSRYLVKRTGEKVFLECVQDMDHENMFWYRQDPGLGL RLIYFSYDVKMKEKGDIPEGYSVSREKKERFSLILESASTNQTSMYLCASSFWGRSTDTQYFGPG TRLTVL (SEQ ID NO:1200). Contrast agents

[0375] In some cases, the TTP of a TMMP comprises a contrast agent or a radiolabel, where the contrast agent facilitates imaging of a tumor to which the TMMP binds.

[0376] Suitable agents include computed tomography (CT), a positron emission tomography (PET), and single photon emission computed tomography (SPECT) radiotracers. Suitable PET / SPECT contrast agents include, e.g., a positron emitter, for example11C,13N,18F,82Ru, and15O. Iodinated CT contrast agents can be used. Suitable contrast agents include gadolinium (Gd), dysprosium, and iron. Gd chelates, such as Gd diethylene triamine pentaacetic acid (GdDTPA), Gd tetraazacyclododecanetetraacetic acid (GdDOTA), polylysine-Gd chelates, and derivatives thereof, can be used. Suitable radioisotopes include123I (iodine),18F (fluorine),99Tc (technetium),111In (indium), and67Ga (gallium). HLA / peptide binding assays

[0377] Whether a given peptide (e.g., a peptide that comprises an epitope) binds a class I HLA (comprising an HLA heavy chain and a β2M polypeptide), and, when bound to the HLA complex, can effectively present an epitope to a TCR, can be determined using any of a number of well-known methods. Assays include binding assays and T-cell activation assays. See, e.g., published PCT Application WO 2020 / 243315 (Cue Biopharma, Inc.). Exemplary TMMPs

[0378] A TMMP comprises at least one heterodimer comprising: a) a first polypeptide comprising: i) a SARS-CoV-2 peptide; and ii) first MHC polypeptide; b) a second polypeptide comprising a second MHC polypeptide, c) at least one immunomodulatory polypeptide, where the first and / or the second polypeptide comprises the immunomodulatory polypeptide; d) an Ig Fc polypeptide ora non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and e) a tumor-targeting polypeptide (TTP), where the first and / or the second polypeptide comprises the TTP. These components can be arranged in any of a variety of configurations. See, e.g., FIG.18A-18C and FIG.28A-28C for non-limiting examples of such configurations.

[0379] Thus, in some cases, a TMMP comprises at least one heterodimer comprising: a) a first polypeptide comprising: i) a SARS-CoV-2 peptide; ii) first MHC class I polypeptide; and iii) at least one immunomodulatory polypeptide; b) a second polypeptide comprising a second MHC class I polypeptide; c) an Ig Fc polypeptide or a non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and d) a TTP, where the first and / or the second polypeptide comprises the TTP. In other instances, a TMMP comprises at least one heterodimer comprising: a) a first polypeptide comprising: i) a SARS-CoV-2 peptide; and ii) first MHC class I polypeptide; b) a second polypeptide comprising: i) a second MHC class I polypeptide; and ii) at least one immunomodulatory polypeptide; c) an Ig Fc polypeptide or a non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and d) a TTP, where the first and / or the second polypeptide comprises the TTP. In some cases, a TMMP comprises at least one heterodimer comprising: a) a first polypeptide comprising: i) a SARS-CoV-2 peptide; ii) first MHC class I polypeptide; and iii) at least one immunomodulatory polypeptide; b) a second polypeptide comprising: i) a second MHC class I polypeptide; and ii) at least one immunomodulatory polypeptide; c) an Ig Fc polypeptide or a non-Ig scaffold, where the first and / or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and d) a TTP, where the first and / or the second polypeptide comprises the TTP. In some cases, the at least one immunomodulatory polypeptide is a wild-type immunomodulatory polypeptide. In other cases, the at least one immunomodulatory polypeptide is a variant immunomodulatory polypeptide that exhibits reduced affinity for a co-immunomodulatory polypeptide, compared to the affinity of a corresponding wild-type immunomodulatory polypeptide for the co-immunomodulatory polypeptide. In some cases, a TMMP comprises two immunomodulatory polypeptides, where the two immunomodulatory polypeptides have the same amino acid sequence. In some cases, the TTP is an antibody specific for a cancer-associated antigen, e.g., a cancer-associated antigen present on the surface of a cancer cell. In some cases, the TTP is a scFv or a nanobody. In some cases, the TTP is an antibody specific for a cancer-associated peptide / HLA complex (i.e., an HLA heavy chain and a β2M polypeptide) present on the surface of a cancer cell. In some cases, the TTP is a single-chain T-cell receptor (scTCR) specific for a cancer-associated antigen, e.g., a cancer-associated antigen present on the surface of a cancer cell.

[0380] For example, in some cases, a TMMP comprises: a) a first polypeptide comprising, in order from N-terminus to C-terminus: i) a SARS-CoV-2 peptide; ii) an optional linker; and iii) a β2M polypeptide; and b) a second polypeptide comprising, in order from N-terminus to C-terminus: i) a TTP;ii) an optional linker; iii) an MHC class I heavy chain polypeptide; iv) an optional linker; v) an Ig Fc polypeptide; vi) an optional linker; and vii) a MOD. In some cases, a TMMP comprises: a) a first polypeptide comprising, in order from N-terminus to C-terminus: i) a SARS-CoV-2 peptide; ii) a linker; and iii) a β2M polypeptide; and b) a second polypeptide comprising, in order from N-terminus to C- terminus: i) a TTP; ii) a linker; iii) an MHC class I heavy chain polypeptide; iv) a linker; v) an Ig Fc polypeptide; vi) a linker; and vii) a MOD. In some cases, a TMMP comprises: a) a first polypeptide comprising, in order from N-terminus to C-terminus: i) a SARS-CoV-2 peptide; ii) a linker; and iii) a β2M polypeptide; and b) a second polypeptide comprising, in order from N-terminus to C-terminus: i) a TTP; ii) a linker; iii) an MHC class I heavy chain polypeptide; iv) a linker; v) an Ig Fc polypeptide; vi) a linker; vii) a first MOD; viii) a linker; and ix) a second MOD. In some cases, the HLA heavy chain comprises an amino acid sequence having at least 95% amino acid sequence identity to an HLA-A*0201, an HLA-A*1101, an HLA-A*2402, or an HLA-A*3303 amino acid sequence depicted in any one of FIG.3A-3E, FIG.4A-4E, FIG.5A-5E, and FIG.6A-6E. In some cases, the HLA heavy chain comprises an amino acid sequence having at least 95% amino acid sequence identity to an HLA-E amino acid sequence depicted in any one of FIG.16A-16D. In some cases, the HLA heavy chain comprises an amino acid sequence having at least 95% amino acid sequence identity to an HLA-G amino acid sequence depicted in any one of FIG.17A-17D. In some cases, the HLA heavy chain polypeptide is an HLA-A*0201 polypeptide. In some cases, the HLA heavy chain polypeptide is an HLA-A*2402 polypeptide. In some cases, the HLA heavy chain polypeptide is an HLA-A*1101 polypeptide. In some cases, the HLA heavy chain polypeptide is an HLA-A*3303 polypeptide. In some cases, the HLA heavy chain polypeptide is an HLA-E polypeptide. In some cases, the HLA heavy chain polypeptide is an HLA-G polypeptide. In some cases, the Ig Fc polypeptide is a human IgG1 Fc polypeptide. In some cases, the Ig Fc polypeptide is an IgG1 Fc polypeptide comprising L234A and L235A substitutions. In some cases, the first and the second polypeptides are disulfide linked to one another. In some cases, the first and the second polypeptides are linked to one another by 2 disulfide bonds. In some cases, the immunomodulatory polypeptide comprises a wild-type amino acid sequence; in other cases, the immunomodulatory polypeptide is a variant, e.g., as described above. In some cases, the immunomodulatory polypeptide is a 4-1BBL polypeptide, e.g., a variant 4-1BBL polypeptide. In some cases, the immunomodulatory polypeptide is a variant IL-2 polypeptide. In some cases, the immunomodulatory polypeptide is a variant IL-2 polypeptide comprising H16A and F42A substitutions. In some cases, the immunomodulatory polypeptide is a variant IL-2 polypeptide comprising H16T and F42A substitutions. In some cases, the second polypeptide comprises a linker between the TTP and the MHC class I heavy chain, where the linker has a length of less than about 25 amino acids, less than about 20 amino acids, less than about 15 amino acids, less than about 10 amino acids, less than about 8 amino acids, or less than about 6 amino acids. In some cases, the second polypeptide comprises a rigid peptidelinker between the Ig Fc polypeptide and the MOD. In some cases, the TTP is an antibody specific for a cancer-associated antigen, e.g., a cancer-associated antigen present on the surface of a cancer cell. In some cases, the TTP is a scFv or a nanobody. In some cases, the TTP is an antibody specific for a cancer-associated peptide / HLA complex (i.e., an HLA heavy chain and a β2M polypeptide) present on the surface of a cancer cell. In some cases, the TTP is a scTCR specific for a cancer-associated antigen, e.g., a cancer-associated antigen present on the surface of a cancer cell. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence YLQPRTFLL (SEQ ID NO:218) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide has an amino acid sequence depicted in FIG.19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0381] In some cases, a TMMP comprises an HLA-A*0201 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the HLA-A*0201 polypeptide depicted in FIG.20B. In some cases, a TMMP comprises an IgG1 Fc polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IgG1 Fc polypeptide depicted in FIG.20B. In some cases, a TMMP comprises an IL-2 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IL-2 polypeptide depicted in FIG.20B.

[0382] In some cases, a TMMP comprises an HLA-A*1101 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the HLA-A*1101 polypeptide depicted in FIG.20D. In some cases, a TMMP comprises an IgG1 Fc polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IgG1 Fc polypeptide depicted in FIG.20D. In some cases, a TMMP comprises an IL-2 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IL-2 polypeptide depicted in FIG.20D.

[0383] In some cases, a TMMP comprises an HLA-A*2402 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the HLA-A*2402 polypeptide depicted in FIG.20F. In some cases, a TMMP comprises an IgG1 Fcpolypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IgG1 Fc polypeptide depicted in FIG.20F. In some cases, a TMMP comprises an IL-2 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IL-2 polypeptide depicted in FIG.20F.

[0384] In some cases, a TMMP comprises an HLA-E*0101 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the HLA-E*0101 polypeptide depicted in FIG.20H. In some cases, a TMMP comprises an IgG1 Fc polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IgG1 Fc polypeptide depicted in FIG.20H. In some cases, a TMMP comprises an IL-2 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IL-2 polypeptide depicted in FIG.20H.

[0385] In some cases, a TMMP comprises an HLA-E*0103 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the HLA-E*0103 polypeptide depicted in FIG.20J. In some cases, a TMMP comprises an IgG1 Fc polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IgG1 Fc polypeptide depicted in FIG.20J. In some cases, a TMMP comprises an IL-2 polypeptide having an amino acid sequence having at least at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the IL-2 polypeptide depicted in FIG.20J.

[0386] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20A; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21A. As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG. 19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ IDNO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0387] In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-Her2 scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG.19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0388] In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-CD22 scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG.19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0389] In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-TROP-2 scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG. 19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0390] In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-mesothelin scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG. 19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0391] In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20A, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-BCMA scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.21A. In some cases, (Xn) is a SARS-CoV-2 peptide having one of the amino acid sequences depicted in FIG. 19. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence GTHWFVTQR (SEQ ID NO:332) and has a length of 9 amino acids. In some cases, the SARS-CoV-2 peptide comprises the amino acid sequence QYIKWPWYI (SEQ ID NO:400) and has a length of 9 amino acids.

[0392] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20A; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21B. In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21B.

[0393] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20A; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21C. In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21C.

[0394] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20A; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21D. In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20A; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21D.

[0395] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20E; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21B. In some cases, a TMMPcomprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20E; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21B.

[0396] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20D; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21C. In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20C; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21C.

[0397] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG.20E; and b) a second polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, or at least 99%, amino acid sequence identity to the amino acid sequence depicted in FIG. 21D. In some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20E; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21D.

[0398] As noted above, in some cases, a TMMP comprises an HLA-E heavy chain polypeptide. In some cases, the TMMP comprises a SARS-CoV-2 peptide having the amino acid sequence YLQPRTLFL (SEQ ID NO:887). In some cases, the TMMP comprises a SARS-CoV-2 peptide having the amino acid sequence YLQPRTLIL (SEQ ID NO:1201). In some cases, the TMMP comprises a SARS-CoV-2 peptide having the amino acid sequence YLQPRTLVL (SEQ ID NO:1202). As one non- limiting example, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20G; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21E. As another non-limiting example, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20I; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.21E. In some cases, the TMMP has a TTP other than an anti-CD19 scFv. For example, in some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 20G, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti- mesothelin scFv, an anti-CD22 scFv, or any other anti-cancer-associated scFv or nanobody; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 21E. As another example, in some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.20I, except that instead of an anti-CD19 scFv, the first polypeptide comprises an anti-mesothelin scFv; and b) a second polypeptide comprising the amino acid sequence depicted in FIG.21E.

[0399] As one non-limiting example, a TMMP comprises: a) a first polypeptide comprising: i) a SARS-CoV2 peptide that has the amino acid sequence YLQPRTFLL (SEQ ID NO:218); ii) a linkercomprising the amino acid sequence GCGGS(GGGS)2 (SEQ ID NO:1203); and iii) a β2M polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following β2M amino acid sequence: IQRTPKIQVYSCHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLL YYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM (SEQ ID NO:2), where amino acid 12 is a Cys; and b) a second polypeptide comprising: i) an anti-CD19 scFv comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following anti-CD19 scFv amino acid sequence: DIQLTQSPASLAVSLGQRATISCKASQSVDYDGDSYLNWYQQIPGQPPKLLIYDASNLVSGIPPRF SGSGSGTDFTLNIHPVEKVDAATYHCQQSTEDPWTFGGGTKLEIKGGGGSGGGGSGGGGSQVQ LQQSGAELVRPGSSVKISCKASGYAFSSYWMNWVKQRPGQGLEWIGQIWPGDGDTNYNGKFK GKATLTADESSSTAYMQLSSLASEDSAVYFCARRETTTVGRYYYAMDYWGQGTTVTVSS (SEQ ID NO:872); ii) a linker, e.g., where the linker has the sequence GGGGS (SEQ ID NO:873); iii) an HLA-A*0201 polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following HLA-A*0201 amino acid sequence: GSHSMRYFFTSVSRPGRGEPRFIAVGYVDDTQFVRFDSDAASQRMEPRAPWIEQEGPEYWDGET RKVKAHSQTHRVDLGTLRGCYNQSEAGSHTVQRMYGCDVGSDWRFLRGYHQYAYDGKDYIA LKEDLRSWTAADMAAQTTKHKWEAAHVAEQLRAYLEGTCVEWLRRYLENGKETLQRTDAPK THMTHHAVSDHEATLRCWALSFYPAEITLTWQRDGEDQTQDTELVETRPCGDGTFQKWAAVV VPSGQEQRYTCHVQHEGLPKPLTLRWE (SEQ ID NO:18), where amino acid 84 is a Cys and where amino acid 236 is a Cys; iv) a linker, e.g., where the linker has the sequence AAAGG (SEQ ID NO:874); v) an IgG Fc polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following IgG Fc amino acid sequence: DKTHTCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEV HNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQ VYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLT VDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG (SEQ ID NO:877), where amino acid 14 is an Ala and where amino acid 15 is an Ala; vi) a linker, e.g., where the linker has the amino acid sequence (GGGGS)3 (SEQ ID NO:875); vii) a first MOD, where the first MOD comprises an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following variant IL-2 amino acid sequence: APTSSSTKKTQLQLEALLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELK PLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIIST LT (SEQ ID NO:878), e.g., where amino acid 16 is an Ala and where amino acid 42 is an Ala; viii) a linker, e.g., where the linker has the amino acid sequence (GGGGS)4 (SEQ ID NO:876); and ix) asecond MOD, where the second MOD comprises an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following variant IL-2 amino acid sequence: APTSSSTKKTQLQLEALLLDLQMILNGINNYKNPKLTRMLTAKFYMPKKATELKHLQCLEEELK PLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIIST LT (SEQ ID NO:878), e.g., where amino acid 16 is an Ala and where amino acid 42 is an Ala. As an example, in some cases, a TMMP comprises a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4518” construct depicted in FIG.32A. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4518” construct depicted in FIG.32A.

[0400] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4728” construct depicted in FIG.38A. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4728” construct depicted in FIG.38A.

[0401] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4730” construct depicted in FIG.38B. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4730” construct depicted in FIG.38B.

[0402] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4701” construct depicted in FIG.38C. In somecases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4701” construct depicted in FIG.38C.

[0403] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4729” construct depicted in FIG.38D. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4729” construct depicted in FIG.38D.

[0404] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4731” construct depicted in FIG.38E. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4731” construct depicted in FIG.38E.

[0405] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4776” construct depicted in FIG.38F. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4776” construct depicted in FIG.38F.

[0406] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4777” construct depicted in FIG.38G. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4777” construct depicted in FIG.38G.

[0407] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4778” construct depicted in FIG.38H. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4778” construct depicted in FIG.38H.

[0408] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4519” construct depicted in FIG.42A. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4519” construct depicted in FIG.42A.

[0409] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4520” construct depicted in FIG.42B. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4520” construct depicted in FIG.42B.

[0410] In some cases, a TMMP comprises: a) a first polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the “4337” construct depicted in FIG.42C. In some cases, a TMMP comprises: a) a first polypeptide having the amino acid sequence of the “4700” construct depicted in FIG.32B; and b) a second polypeptide having the amino acid sequence of the “4337” construct depicted in FIG.42C.

[0411] In some cases, a TMMP comprises: a) ) a first polypeptide comprising: i) a peptide epitope having a length of from 4 amino acids to 25 amino acids (e.g., from 6 to 12 amino acids; from 8to 12 amino acids; etc.); ii) a linker having the amino acid sequence GCGGS(GGGGS)2 (SEQ ID NO:882); and iii) a β2M polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the β2M amino acid sequence depicted in FIG.32G; and b) a second polypeptide of the structure: [X]-GGGGS-[Y]-AAAGG-[Z]-(GGGGS)3- APTSSSTKKTQLQLEX1LLLDLQMILNGINNYKNPKLTRMLTX2KFYMPKKATELKHLQCLEEEL KPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIIS TLT-(GGGGS)4- APTSSSTKKTQLQLEX1LLLDLQMILNGINNYKNPKLTRMLTX2KFYMPKKATELKHLQCLEEEL KPLEEVLNLAQSKNFHLRPRDLISNINVIVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIIS TLT (SEQ ID NO:1217), where [X] is a scFv specific for a cancer-associated antigen or a nanobody specific for a cancer-associated antigen; where [Y] is an HLA heavy chain polypeptide; where [Z] is an Ig Fc polypeptide; where X1 is an amino acid other than His; and where X2 is an amino acid other than Phe. Such a TMMP is depicted in FIG.40A and FIG. 40B. In some cases, the Ig Fc polypeptide is a variant of a Fc polypeptide such as a human IgG1 Fc, which variant has a substantially reduced ability to effect CDC or ADCC.

[0412] In some cases, a TMMP comprises: a) a first polypeptide comprising: i) a peptide epitope having a length of from 4 amino acids to 25 amino acids (e.g., from 6 to 12 amino acids; from 8 to 12 amino acids; etc.); ii) a linker having the amino acid sequence GCGGS(GGGGS)2 (SEQ ID NO:882); and iii) a β2M polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the β2M amino acid sequence depicted in FIG.32G; and b) a second polypeptide comprising: i) a scFv polypeptide or a nanobody specific for a cancer-associated antigen; ii) a linker having the sequence GGGGS (SEQ ID NO:873); iii) an HLA heavy chain polypeptide; iv) a linker having the sequence AAAGG (SEQ ID NO:874); v) an Ig Fc polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the Ig Fc amino acid sequence depicted in FIG.38K; vi) a linker having the sequence (GGGGS)3 (SEQ ID NO:875); vii) a variant IL-2 polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the variant IL-2 amino acid sequence depicted in FIG.38L, where amino acid 16 is other than His (e.g., where amino acid 16 is Ala) and where amino acid 42 is other than Phe (e.g., where amino acid 42 is Ala); viii) a linker having the sequence (GGGGS)4 (SEQ ID NO:876); and ix) a variant IL-2 polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the variant IL-2 amino acid sequence depicted in FIG.38L, where amino acid 16 is other than His (e.g., where amino acid 16 is Ala) and where amino acid 42 is other than Phe (e.g., where amino acid 42 is Ala). Such a TMMP is depicted in FIG.40A and FIG.40B. In some cases, the Ig Fc polypeptide is a variant of a Fc polypeptide such as a human IgG1 Fc, which variant has a substantiallyreduced ability to effect CDC or ADCC; for example, in some cases, the variant Ig Fc polypeptide comprises an Ala (instead of a Leu) at position 14 and an Ala (instead of a Leu) at position 15, based on the amino acid numbering set out in FIG. 38K. In some cases, the peptide epitope is a SARS-CoV-2 peptide. In some cases, the HLA heavy chain polypeptide is an HLA-A*0201 polypeptide; e.g., an HLA polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG. 3A. In some cases, the HLA heavy chain polypeptide is an HLA-A*0201 polypeptide comprising Y84C and A236C substitutions (e.g., as depicted in FIG.3C). In some cases, the HLA heavy chain polypeptide is an HLA- A*1101 polypeptide; e.g., an HLA polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.4A. In some cases, the HLA heavy chain polypeptide is an HLA-A*1101 polypeptide comprising Y84C and A236C substitutions (e.g., as depicted in FIG.4C). In some cases, the HLA heavy chain polypeptide is an HLA-A*2402 polypeptide; e.g., an HLA polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.5A. In some cases, the HLA heavy chain polypeptide is an HLA- A*2402 polypeptide comprising Y84C and A236C substitutions (e.g., as depicted in FIG.5C). In some cases, the HLA heavy chain polypeptide is an HLA-A*3303 polypeptide; e.g., an HLA polypeptide comprising an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence depicted in FIG.6A. In some cases, the HLA heavy chain polypeptide is an HLA-A*3303 polypeptide comprising Y84C and A236C substitutions (e.g., as depicted in FIG. 6C). In some cases, the HLA heavy chain polypeptide is an HLA-E polypeptide. In some cases, the HLA-E polypeptide comprises an amino acid sequence having at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the HLA-E amino acid sequence depicted in any one of FIG.16A-16D. In some cases, the HLA-E polypeptide comprises Y84C and A236C substitutions (e.g., as depicted in FIG.16B and FIG.16D). In some cases, the HLA heavy chain polypeptide is an HLA-E polypeptide, and the peptide epitope is a SARS-CoV2 peptide having the amino acid sequence VMPLSAPTL (SEQ ID NO:914). In some cases, the HLA heavy chain polypeptide is an HLA-E polypeptide, and the peptide epitope is a SARS-CoV2 peptide having the amino acid sequence YLQPRTFLL (SEQ ID NO:218). In some cases, the scFv polypeptide or the nanobody specifically binds CD19. In some cases, the scFv polypeptide or the nanobody specifically binds mesothelin. In some cases, the scFv polypeptide specifically binds TROP-2. In some cases, the scFv polypeptide or the nanobody specifically binds CD28. In some cases, the scFv polypeptide or the nanobody specifically binds Her2. In some cases, the scFv polypeptide or the nanobody specifically binds PSMA. In some cases, the scFv polypeptide or the nanobody specifically binds CD22. In some cases, the scFv polypeptide or the nanobody specifically binds BCMA. In some cases, the scFv polypeptide or thenanobody specifically binds MUC1. In some cases, the scFv polypeptide or the nanobody specifically binds MUC16. In some cases, the scFv polypeptide or the nanobody specifically binds claudin-18.2. In some cases, the TMMP comprises a scFv. As one non-limiting example, in some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG. 41B; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 41A. As another non-limiting example, in some cases, a TMMP comprises: a) a first polypeptide comprising the amino acid sequence depicted in FIG.41C; and b) a second polypeptide comprising the amino acid sequence depicted in FIG. 41A.

[0413] Although embodiments including a SARS-CoV-2 are illustrated herein, as discussed above, peptide epitopes from other pathogens could be employed in TMMPs that also comprise one or more MODs. For example, other suitable epitopes include, but are not limited to, epitopes present in an infectious disease agent, e.g., an epitope presented by a virus-encoded polypeptide or bacteria. Examples of viral infectious disease agents include, e.g., Adenoviruses, Adeno-associated virus, Alphaviruses (Togaviruses), Eastern equine encephalitis virus, Eastern equine encephalomyelitis virus, Venezuelan equine encephalomyelitis vaccine strain TC-83, Western equine encephalomyelitis virus, Arenaviruses, Lymphocytic choriomeningitis virus (non-neurotropic strains), Tacaribe virus complex, Bunyaviruses, Bunyamwera virus, Rift Valley fever virus vaccine strain MP-12, Chikungunya virus, Calciviruses, Coronaviruses, Cowpox virus, Flaviviruses (Togaviruses)-Group B Arboviruses, Dengue virus serotypes 1, 2, 3, and 4, Yellow fever virus vaccine strain 17D, Hepatitis A, B, C, D, and E viruses, the Cytomegalovirus, Epstein Barr virus, Eastern Equine encephalitis virus, Herpes simplex types 1 and 2, Herpes zoster, Human herpesvirus types 6 and 7, hepatitis C virus (HVC), hepatitis B virus (HBV), Influenza viruses types A, B, and C, Papovaviruses, Newcastle disease virus, Measles virus, Mumps virus, Parainfluenza viruses types 1, 2, 3, and 4, polyomaviruses (JC virus, BK virus), Respiratory syncytial virus, Human parvovirus (B 19), Coxsackie viruses types A and B, Echoviruses, Polioviruses, Rhinoviruses, Alastrim (Variola minor virus), Smallpox (Variola major virus), Whitepox Reoviruses, Coltivirus, human Rotavirus, and Orbivirus (Colorado tick fever virus), Rabies virus, Vesicular stomatitis virus, Rubivirus (rubella), Semliki Forest virus, St. Louis encephalitis virus, Venezuelan equine encephalitis virus, Venezuelan equine encephalomyelitis virus, Arenaviruses (a.k.a. South American Hemorrhagic Fever virus), Flexal, Lymphocytic choriomeningitis virus (LCM) (neurotropic strains), Hantaviruses including Hantaan virus, Rift Valley fever virus, Japanese encephalitis virus, Yellow fever virus, Monkeypox virus, Human immunodeficiency virus (HIV) types 1 and 2, Human T cell lymphotropic virus (HTLV) types 1 and 2, Simian immunodeficiency virus (SIV), Vesicular stomatitis virus, Guanarito virus, Lassa fever virus, Junin virus, Machupo virus, Sabia, Crimean-Congo hemorrhagic fever virus, Ebola viruses, Marburg virus, Tick-borne encephalitis virus complex (flavi) including Central European tick-borne encephalitis, Far Eastern tick-borne encephalitis, Hanzalova,Hypr, Kumlinge, Kyasanur Forest disease, Omsk hemorrhagic fever, and Russian Spring Summer encephalitis viruses,Herpesvirus simiae (Herpes B or Monkey B virus), Cercopithecine herpesvirus 1 (Herpes B virus), Equine morbillivirus (Hendra and Hendra-like viruses), Nipah virus, Variola major virus (Smallpox virus), Variola minor virus (Alastrim), African swine fever virus, African horse sickness virus, Akabane virus, Avian influenza virus (highly pathogenic), Blue tongue virus, Camel pox virus, Classical swine fever virus, Cowdria ruminantium (heartwater), Foot and mouth disease virus, Goat pox virus, Japanese encephalitis virus, Lumpy skin disease virus, Malignant catarrhal fever virus, Menangle virus, Newcastle disease virus (VVND), Vesicular stomatitis virus (exotic), and Zika virus. Antigens encoded by such viruses are known in the art; a peptide epitope suitable for use in a TMMP of the present disclosure can include a peptide from any known viral antigen. In some cases, a viral epitope is an epitope present in a viral antigen encoded by a virus other than SARS-CoV-2 that infects a majority of the human population, where such viruses include, e.g., cytomegalovirus (CMV), Epstein-Barr virus (EBV), human papilloma virus, adenovirus, and the like. See, e.g., published PCT application WO2020 / 243315 (Cue Biopharma, Inc.) at paragraphs

[0321] -

[0338] . Accordingly, this disclosure expressly contemplates TMMPs comprising an epitope other than a SARS-CoV-2 epitope. TMMPS WITHOUT ANIMMUNOMODULATORYPOLYPEPTIDE

[0414] The present disclosure provides a TMMP comprising: a) a first polypeptide; and b) a second polypeptide, wherein the TMMP comprises: a peptide epitope; a first MHC polypeptide; a second MHC polypeptide; an immunoglobulin (Ig) Fc polypeptide or a non-Ig scaffold; and a TTP, where the TMMP does not include an immunomodulatory polypeptide (e.g., does not comprise a MOD as set forth in Table 2; e.g., does not comprise an IL-2 polypeptide, a 4-1BBL polypeptide, etc.). Such a TMMP may also be referred to herein as a “MODless TMMP.” The components of a MODless TMMP (e.g., peptide epitope, first MHC polypeptide, second MHC polypeptide, Ig Fc polypeptide or non-Ig scaffold, and TTP) can be arranged in a variety of configurations. The components (other than MODs) are as described above.

[0415] In some cases, the present disclosure provides a protein comprising at least one heterodimer comprising: a) a first polypeptide comprising: i) a SARS-CoV-2 peptide, wherein the SARS-CoV-2 peptide has a length of from about 4 amino acids to about 25 amino acids, e.g., from 8 to 12 amino acids; and a β2M polypeptide; b) a second polypeptide an MHC class I heavy chain polypeptide; c) an Ig Fc polypeptide or a non-Ig scaffold, where the first or the second polypeptide comprises the Ig Fc polypeptide or the non-Ig scaffold; and d) a TTP, where the first and / or the second polypeptide comprises the TTP, wherein the protein does not include an immunomodulatory polypeptide. One or more of the individual components of the first and / or second polypeptide can be linked together by one or more linkers. Suitable SARS-CoV-2 peptides, MHC class I heavy chain polypeptides, and Ig Fc polypeptides are described above. Such a protein can also be referred to as a “MOD-less TMMP.”

[0416] In some cases, a MOD-less TMMP is dimerized. Thus, present disclosure thus provides a protein that is a dimerized MOD-less TMMP comprising two MOD-less TMMPs that are covalently linked to each other. The covalent linkage of the dimer can be one or more disulfide bonds between an Ig Fc polypeptide in the first MOD-less TMMP and an Ig Fc polypeptide in the second MOD-less TMMP.

[0417] In some cases, a MODless TMMP comprises: a) a first polypeptide comprising: (i) a peptide epitope, where the peptide epitope is a SARS-CoV-2 peptide; and (ii) a first MHC polypeptide, where the first MHC polypeptide is a β2M polypeptide; and b) a second polypeptide comprising: (i) a TTP; (ii) a second MHC polypeptide, where the second MHC polypeptide is an MHC class I heavy chain polypeptide; and (iii) an Ig Fc polypeptide. A peptide linker may be interposed between one or more of: (i) the peptide epitope and the β2M polypeptide; (ii) the MHC class I heavy chain polypeptide and the TTP; (iii) the MHC class I heavy chain polypeptide and the Ig Fc polypeptide. In some cases, the first polypeptide comprises, in order from N-terminus to C-terminus: (i) a SARS-CoV-2 peptide; and (ii) a β2M polypeptide. In some cases, the first polypeptide comprises, in order from N-terminus to C- terminus: (i) a SARS-CoV-2 peptide; (ii) a peptide linker, where the peptide linker optionally may include a Cys residue; and (iii) a β2M polypeptide. In some cases, the second polypeptide comprises, in order from N-terminus to C-terminus: (i) a TTP; (ii) an optional linker; (iii) an MHC class I heavy chain polypeptide; (iv) an optional linker; and (v) an Ig Fc polypeptide. In some cases, the second polypeptide comprises, in order from N-terminus to C-terminus: (i) an MHC class I heavy chain polypeptide; (ii) an optional linker; (iii) an Ig Fc polypeptide; (iv) an optional linker; and (v) a TTP. In some cases, the second polypeptide comprises, in order from N-terminus to C-terminus: (i) an MHC class I heavy chain polypeptide; (ii) an optional linker; (iii) a TTP; (iv) an optional linker; and (v) an Ig Fc polypeptide. In some cases, the β2M polypeptide comprises a Cys residue, e.g., a Cys residue at position 12 according to the amino acid numbering of the β2M amino acid sequence depicted in FIG.1B. In some cases, the MHC class I heavy chain polypeptide comprises a Cys residue at position 84 and / or a Cys residue at position 236.

[0418] The following are non-limiting examples of MODless TMMPs.

[0419] In some cases, a MODless TMMP comprises: a) a first polypeptide comprising: (i) a SARS-CoV-2 peptide; and (ii) a β2M polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following β2M amino acid sequence: IQRTPKIQVYSCHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLL YYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM (SEQ ID NO:2), where amino acid 12 is a Cys; and b) a second polypeptide comprising: (i) an anti-CD19 scFv polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequenceidentity to the anti-CD19 scFv polypeptide amino acid sequence depicted in FIG.32C; (ii) an MHC class I heavy chain polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the HLA-A*0201 (Y84C; A236C) amino acid sequence depicted in FIG.32D, where amino acid 84 is Cys and where amino acid 236 is Cys; and (iii) an Ig Fc polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the Ig Fc (L14A; L15A) amino acid sequence depicted in FIG.32E, where amino acid 14 is an Ala and where amino acid 15 is an Ala. In some cases, the first polypeptide comprises a Cys-containing peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide. As an example, in some cases, the first polypeptide comprises, between the SARS-CoV-2 peptide and the β2M polypeptide, a peptide linker of the following amino acid sequence: GCGGSGGGGSGGGGS (SEQ ID NO:882). In some cases, the SARS-CoV-2 peptide has the amino acid sequence YLQPRTFLL (SEQ ID NO:218). In some cases, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the polypeptide designated “4700” and depicted in FIG. 32B. In some cases, the first polypeptide comprises an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the amino acid sequence of the polypeptide designated “4656” and depicted in FIG.33.

[0420] In some cases, a MODless TMMP comprises: a) a first polypeptide comprising: (i) a SARS-CoV-2 peptide; and (ii) a β2M polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the following β2M amino acid sequence: IQRTPKIQVYSCHPAENGKSNFLNCYVSGFHPSDIEVDLLKNGERIEKVEHSDLSFSKDWSFYLL YYTEFTPTEKDEYACRVNHVTLSQPKIVKWDRDM (SEQ ID NO:2), where amino acid 12 is a Cys; and b) a second polypeptide comprising: (i) an anti-CD19 scFv polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the anti-CD19 scFv polypeptide amino acid sequence depicted in FIG.32C; (ii) an MHC class I heavy chain polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the HLA-E (Y84C; A236C) amino acid sequence depicted in FIG.16B or FIG.16D, where amino acid 84 is Cys and where amino acid 236 is Cys; and (iii) an Ig Fc polypeptide comprising an amino acid sequence having at least 90%, at least 95%, at least 98%, at least 99%, or 100%, amino acid sequence identity to the Ig Fc (L14A; L15A) amino acid sequence depicted in FIG.32E, where amino acid 14 is an Ala and where amino acid 15 is an Ala. In some cases, the first polypeptide comprises a Cys-containing peptide linker between the SARS-CoV-2 peptide and the β2M polypeptide. As an example, in some cases, the first polypeptide comprises, between the SARS-CoV-2 peptide and the β2M polypeptide, a peptide linker of the following amino acidsequence: GCGGSGGGGSGGGGS (SEQ ID NO:882). In some cases, the SARS-CoV-2 peptide has the amino acid sequence VMPLSAPTL (SEQ ID NO:914).

[0421] Although MODless embodiments including a SARS-CoV-2 are illustrated herein, those skilled in the art will readily understand that other peptide epitopes could be employed in MODless TMMPs. For example, other suitable epitopes include, but are not limited to, epitopes present in an infectious disease agent, e.g., an epitope presented by a virus-encoded polypeptide. Examples of viral infectious disease agents include, e.g., Adenoviruses, Adeno-associated virus, Alphaviruses (Togaviruses), Eastern equine encephalitis virus, Eastern equine encephalomyelitis virus, Venezuelan equine encephalomyelitis vaccine strain TC-83, Western equine encephalomyelitis virus, Arenaviruses, Lymphocytic choriomeningitis virus (non-neurotropic strains), Tacaribe virus complex, Bunyaviruses, Bunyamwera virus, Rift Valley fever virus vaccine strain MP-12, Chikungunya virus, Calciviruses, Coronaviruses, Cowpox virus, Flaviviruses (Togaviruses)-Group B Arboviruses, Dengue virus serotypes 1, 2, 3, and 4, Yellow fever virus vaccine strain 17D, Hepatitis A, B, C, D, and E viruses, the Cytomegalovirus, Epstein Barr virus, Eastern Equine encephalitis virus, Herpes simplex types 1 and 2, Herpes zoster, Human herpesvirus types 6 and 7, hepatitis C virus (HVC), hepatitis B virus (HBV), Influenza viruses types A, B, and C, Papovaviruses, Newcastle disease virus, Measles virus, Mumps virus, Parainfluenza viruses types 1, 2, 3, and 4, polyomaviruses (JC virus, BK virus), Respiratory syncytial virus, Human parvovirus (B 19), Coxsackie viruses types A and B, Echoviruses, Polioviruses, Rhinoviruses, Alastrim (Variola minor virus), Smallpox (Variola major virus), Whitepox Reoviruses, Coltivirus, human Rotavirus, and Orbivirus (Colorado tick fever virus), Rabies virus, Vesicular stomatitis virus, Rubivirus (rubella), Semliki Forest virus, St. Louis encephalitis virus, Venezuelan equine encephalitis virus, Venezuelan equine encephalomyelitis virus, Arenaviruses (a.k.a. Sout...

Claims

CLAIMS What is claimed is:

1. A T-cell modulatory multimeric polypeptide (TMMP) comprising at least one heterodimer comprising: a) a first polypeptide comprising, from N-terminus to C-terminus: i) a SARS-CoV-2 peptide, wherein the SARS-CoV-2 peptide has a length of from about 4 amino acids to about 25 amino acids, ii) an optional peptide linker, and iii) first major histocompatibility complex (MHC) polypeptide, wherein the first MHC polypeptide is a β2 microglobulin (β2M) polypeptide; and b) a second polypeptide comprising, from N-terminus to C-terminus: i) a tumor-targeting polypeptide (TTP), ii) a peptide linker, iii) a second MHC polypeptide, wherein the second MHC polypeptide is an MHC class I heavy chain polypeptide, iv) an optional peptide linker, v) an immunoglobulin (Ig) Fc polypeptide, vi) an optional peptide linker, and vii) at least one immunomodulatory polypeptide, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide comprises no more than about 12 amino acids, optionally wherein the peptide linker is no more than 10 amino acids in length (e.g., a (GGGGS)2linker), no more than 8 amino acids in length, no more than 7 amino acids in length, no more than 6 amino acids in length, or no more than 5 amino acids in length, (e.g., a GGGGS linker).

2. The TMMP of claim 1, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide is 5 amino acids in length and has the amino acid sequence GGGGS or is 10 amino acids in length and has the amino acid sequence (GGGGS)2.

3. The TMMP of claim 1, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide is 5 amino acids in length and has the amino acid sequence GGGGS.

4. The TMMP of claim 1, wherein the at least one immunomodulatory polypeptide is a variant IL-2 polypeptide.

5. The TMMP of claim 4, wherein the at least one variant IL-2 polypeptide binds to IL-2R and exhibits decreased or substantially no binding to IL-2Rα, and also exhibits decreased binding to IL- 2Rβ such that the IL-2 variant polypeptide exhibits an overall reduced affinity for IL-2R, optionally wherein the at least one variant comprises substitutions at H16 and F42 selected from H16A, F42A; H16T, F42A; H16E, F42A; H16D,F42A; H16A, F42K; H16T, F42K; H16E, F42K; and H16D, F42K.

6. The TMMP of claim 5, wherein the at least one immunomodulatory polypeptide is a single copy of a variant IL-2 polypeptide.

7. The TMMP of claim 6, wherein the variant IL-2 polypeptide comprises substitutions at H16 and F42.

8. The TMMP of claim 7, wherein the variant IL-2 polypeptide comprises H16A and F42A substitutions.

9. The TMMP of claim 8, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide has the amino acid sequence GGGGS.

10. The TMMP of claim 9, wherein the MHC class I heavy chain polypeptide is an HLA-A02 polypeptide and the SARS-CoV-2 peptide is YLQPRTFLL, or the MHC class I heavy chain polypeptide is an HLA-E polypeptide and the SARS-CoV-2 peptide is VMPLSAPTL or YLQPRTFLL.

11. A homodimer comprising two heterodimers according to any one of claims 1-10, wherein the two heterodimers are covalently linked to one another by one or more disulfide bonds between Ig Fc polypeptides present in the first and second heterodimers.

12. A method of treating a patient having a cancer, the method comprising administering to the patient an effective amount of a pharmaceutical composition comprising an effective amount of a TMMP according to any one of claims 1-10.

13. A method of treating a patient having a cancer, the method comprising administering to the patient an effective amount of a pharmaceutical composition comprising an effective amount of a homodimer comprising two heterodimers according to any one of claims 1-10, wherein the two heterodimers are covalently linked to one another by one or more disulfide bonds between Ig Fc polypeptides present in the first and second heterodimers.

14. A T-cell modulatory multimeric polypeptide (TMMP) comprising at least one heterodimer comprising: a) a first polypeptide comprising i) a SARS-CoV-2 peptide, wherein the SARS-CoV-2 peptide has a length of from about 4 amino acids to about 25 amino acids, ii) an optional peptide linker, and iii) first major histocompatibility complex (MHC) polypeptide, wherein the first MHC polypeptide is a β2 microglobulin (β2M) polypeptide; and b) a second polypeptide comprising, from N-terminus to C-terminus i) a tumor-targeting polypeptide (TTP), ii) a peptide linker, iii) a second MHC polypeptide, wherein the second MHC polypeptide is an MHC class I heavy chain polypeptide, iv) an optional peptide linker, and v) an immunoglobulin (Ig) Fc polypeptide, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide comprises no more than about 12 amino acids, optionally wherein the peptide linker is no more than 10 amino acids in length (e.g., a (GGGGS)2linker), no more than 8 amino acids in length, no more than 7 amino acids in length, no more than 6 amino acids in length, or no more than five amino acids in length, (e.g., a GGGGS linker).

15. The TMMP of claim 14, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide has the amino acid sequence GGGGS or (GGGGS)2.

16. The TMMP of claim 15, wherein the peptide linker between the TTP and the MHC class I heavy chain polypeptide has the amino acid sequence GGGGS.

17. The TMMP of claim 16, wherein:the MHC class I heavy chain polypeptide is an HLA-A02 polypeptide and the SARS-CoV-2 peptide is YLQPRTFLL, or the MHC class I heavy chain polypeptide is an HLA-E polypeptide and the SARS-CoV-2 peptide is VMPLSAPTL or YLQPRTFLL.

18. A homodimer comprising two heterodimers according to any one of claims 14-17, wherein the two heterodimers are covalently linked to one another by one or more disulfide bonds between Ig Fc polypeptides present in the first and second heterodimers.

19. A method of treating a patient having a cancer, the method comprising administering to the patient an effective amount of a pharmaceutical composition comprising an effective amount of a TMMP according to any one of claims 14-17.

20. A method of treating a patient having a cancer, the method comprising administering to the patient an effective amount of a pharmaceutical composition comprising an effective amount of a homodimer comprising two heterodimers according to any one of claims 14-17, wherein the two heterodimers are covalently linked to one another by one or more disulfide bonds between Ig Fc polypeptides present in the first and second heterodimers.

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